issue 1 operator guide - marshall scientific...3.1.6 cytospin ® 3 is a class 1 instrument as...

Thermo Shandon Limited is an ISO 9001 and TickIT Accredited Company.

Iss 1.11CNUTP-GB-EFG

®

74010121 GB

OPERATORGUIDE

ENGLISHIssue 1

CELL PREPARATION SYSTEM

ii1CNUTP-GB-EFG Iss 1.1

© 1997 Thermo Shandon Limited. All rights reserved.

This manual may not, in whole or in part, be copied, photocopied, reproduced, translated, or converted to any electronicor machine readable form without prior written consent of Thermo Shandon Limited.

Thermo Shandon makes every endeavour to ensure that the information contained in its supportdocumentation is correct and clearly stated but does not accept responsibility for any errors or omissions.The development of Thermo Shandon products and services is continuous. Make sure that any publishedinformation that you use for reference is up to date and relates to the status of the product. If necessary,check with Thermo Shandon or your local Thermo Shandon representative.

Safety

Operating Principles; Construction

To Enter; To Start; To Save; To Run; To View; To Repeat; To Stop.

Equipment Checks, Alarms.

Methodology Guidelines

Iss 1.1

iii1CNUTP-GB-EFG

iv1CNUTP-GB-EFG Iss 1.1

Iss 1.1

1.1

1CNU01-GB-EFG

1.1.1 Welcome to the Cytospin ® 3 Cel lPreparation System. Designed and made with care,Cytospin ® is safe to use, simple to operate, andeasy to maintain.

1.1.2 This manual gives instruct ions for i tscorrect operation and use. The number by the sideof each illustration is the same as the paragraphnumber of its associated text.

1.2.1 Thermo Shandon instruments are designedto operate safely and consistently for many years.However, incorrect actions by a user may damagethe equipment, or cause a hazard to health. It isImportant for you to know that:

i The instrument weighs approximately 18Kilograms (39 3/4 lbs); if necessary, get helpto move or lift it.

ii Dangerous voltages are contained inside.Disconnect the instrument from the supply, ortake appropriate precautions, before youundo any panels or remove any accesscovers unless instructed otherwise.

iii Always load and unload the Sealed Head in abiologically safe fume cupboard. Assume thatthe samples are biologically hazardous.

iv Use only genuine Thermo Shandonreplacement parts should a repair becomenecessary.

v Make sure that no fingers or articles ofclothing are allowed within the area definedby the frame assembly if the Cytospin® has tobe operated whi le a panel is removed.Components may suddenly move undercomputer control and cause injury.

vi Cytospin® is not intended for use wi thflammable materials or solvents, nor withmater ia ls which could be explosive orchemically reactive.

vii Bioseals and other biosafety componentscannot be relied upon as a sole safeguardagainst contaminat ion by pathogenicmicro-organisms.

1.2.2 In compliance with International health andsafety guidelines, all our equipment is designed toaccepted standards of safety. Its use does not entailany hazard if operated in accordance with theinstructions given in this manual. However, youmust obey the following safety precautions.

i All users must read and understand theOperator Guide, and only operate the unit inaccordance with the instructions.

ii Potentially lethal voltages above 110V a.c. or50V d.c. are present inside the unit. Do notremove any access covers unless specificallyinstructed to do so by a Thermo Shandonrepresentative.

iii It is important that normal standards of safetyand good laboratory and housekeepingpractices are employed. Always use commonsense when operating the instrument.

iv Any problems should be referred to ourService Department.

v Correct maintenance procedures areessential to ensure safe operation andconsistent performance. It is important thatthe instrument is by serviced by qualifiedThermo Shandon service personnel. Thepurchase of a Maintenance Contract isstrongly recommended.

Iss 1.1

1.2

1CNU01-GB-EFG

THIS SYMBOL APPEARS INDOCUMENTS TO WARN THATINSTRUCTIONS MUST BEFOLLOWED TO ENSURE SAFEAND CORRECT OPERATION.

A warning is g iven in thedocument if there is a danger

of personal injury or damage to equipment.

Note1 Notes give more information about a job

or instruction but do not form part of theinstruction.

Push-button operations performed by the operatorare shown thus: [PUSH-BUTTON TITLE]

This product meets recognised International standards, and complies with all relevantEuropean Directives, only when used with genuine Thermo Shandon consumables,accessories, and spare parts. For further information please refer to the Declaration ofConformity at the back of this publication.

WARNING

®

Cell Preparation System

2.1.1 Cytospin® 3 is designed specifically foruse as a cell preparation system that usescentrifugal force to deposit cells onto a slide formicroscopical analysis.

2.1.2 Specimens are held in up to 12 sampleholders. Each sample holder consists of:

i a stainless steel Cytoclip™ slide clip(1),

ii a microscope slide (2), and

iii disposable Cytofunnel® sample chamber(3) with integral filter card (4)...

OR

separate filter cards with re-usable samplechambers.

2.1.3 The sample holders fit into a SealedHead assembly that is removable so that it canbe loaded and unloaded in a biologically safearea. The sample holders are held t i l ted(Load Position i ) when the instrument isstopped so that the cells in suspension cannottrickle forward and be absorbed by the filtercard.

2.1.4 The sample holders t i l t forward(Operating Position ii) when the Sealed Headrotates so that cel ls in suspension areforcefully deposited out onto a microscopeslide. Excess fluid is absorbed by the filtercard.

Iss 1.1

2.1

1CNU02-GB-EFG

2.1.3

2.1.2

2.1.4

Operating Position ( ii )

Load Position ( i )

1 2 3

4

2.1.5 The speed at which the Sealed Headspins, and the duration, can be programmedwithin the range 200-2000 r.p.m., and 1-99minutes respectively. The acceleration ratesmay be High, Medium or Low.

2.1.6 Up to nine programs can be held in thememory of Cytospin®.

2.2.1 Cytospin® comprises a two part metalhousing. The Sealed Head rotor assembly fitsinto a sprung bowl enclosure in the upper partwhich is closed by a bronzed, see-through,safety cover. The safety cover locks closedduring operation.

2.2.2 A tapered boss in the sprung bowl sup-ports the Sealed Head assembly. Rotarymotion is transmitted to the Sealed Head asthe boss rotates.

2.2.3 The Sealed Head assembly has its ownbronzed see-through lid to seal the cells in sus-pension from the environment.

2.2.4 The lower metal housing encloses theelectrical power and control equipment; and themotor.

2.2.5 Control of the unit is by membranetype, touch sensitive, switches on the frontmounted control panel. Associated displaysshow the selections made.

Iss 1.1

2.2

1CNU02-GB-EFG

2.2.1

2.2.5

2.2.2

Iss 1.1

3.1

3.1.1 Cytospin® 3 is a precision instrumentthat must be unpacked and installed with care.Make sure that the detai l on the cartoncorresponds with the Purchase Order beforethe Cytospin is removed from its carton.

3.1.2 Make sure that the bench or supportingsurface on which Cytospin® is to operate is asolid immovable surface that is both flat andlevel. There must be a clear space aroundCytospin®, of at least 300mm (12 in.), that isfree of hazardous materials, debris, orpersonnel. The bench must be stable and capa-ble of supporting a weight of 18 kg (39 3/4lb), and the environment must be dust free.

Note1 Do not place any paper or protective

sheet between the Cytospin® and thesupporting surface or the relative motoncut-out may not operate.

3.1.3 If necessary, get someone to help youlift the carton; it weighs approximately 20 kg(44lb). Grip the underneath of the carton firmlyto move or lift it. Remove the outer packing.Check that the instrument is not damaged, thengrip the underside edges of the upper body andlift the Cytospin out of the carton. Read thelabel at the rear of the instrument and makesure that the power supply requirements arecorrect.3.1.4 Undo the accessories pack. Check that

1CNU03-GB-EFG

3.1.3

3.1.2

Product:Cat No:Serial No:

Nominal Ratings:

CYTOSPIN 374000201MA631510W

220 - 240 V~50/60 Hz150VA

MADE IN UK

Astmoor, Runcorn, Cheshire, WA7 1PR, UKTHERMO SHANDON LIMITED

®

PATENTS: EP0047840 US: 4391710 JAP: TOKKYO 1431078

Product:Cat No:Serial No:

Nominal Ratings:

CYTOSPIN 374000222 Issue 7MA2256A0002

110 - 120 V~50/60 Hz150VA

MADE IN UK

Astmoor, Runcorn, Cheshire, WA7 1PR, UKTHERMO SHANDON LIMITED

®

PATENTS: EP0047840 US: 4391710 JAP: TOKKYO 1431076

®

NRTL/C

the items are present as listed.

Note1 Inform your Thermo Shandon dealer

immediately if there are any breakages orshortages. Quote the number of yourOrder, the Invoice, the Delivery Note and itsdate.

2 Important information with respect to Parts isprinted on the pack in which Cytospin® issupplied. Keep this information in a safeplace.

3 The ~ symbol on the rating plate indicatesthat the instrument operates on AlternatingCurrent supplies (a.c.)

Injury or damage may result ifinstructions 3.1.5 and 3.1.6

are not followed

3.1.5 Select the appropriate mains lead andfit it into the fused mains input socket at therear of the unit. Only mains leads less than 3mlong should be used with this instrument.

Notes1 Mains input fuses in instruments that operate

from a 110V supply are T2.0 Amp.

2 Mains input fuses in instruments that operatefrom a 240V supply are T0.8 Amp.

3 On 240V instruments the mains supply plugshould be fitted with a 5 Amp fuse.

4 Only a technically competent person shouldbe employed to change fuses.

3.1.6 Cytospin® 3 is a Class 1 instrument as

Iss 1.11CNU03-GB-EFG

3.2

WARNUNG

defined in IEC1010 and must be protectivelyearthed. Make sure that the instrument isproperly connected to a good Earth/Groundcontact, marked as shown.

3.1.7 In an emergency it must be possible tointerrupt the mains supply from a place eithernear an exit that is remote from the Cytospin®,or outside the room. Interruption of the supplycan be by switching off at the supply socket, orby removing the plug.

3.1.8 Press O of the ON/OFF ( l / O ) switchon the front of the instrument to ensure that theinstrument is switched off.

3.1.9 Plug the mains cable into the powersupply, and switch on the power supply.

3.1.10 Press l of the ON/OFF ( l / O ) switchto switch the instrument ON.

3.1.11 If the ' BAL' and ' ! ' alarm lights flash,lift the front of the instrument approximately10mm (½ in.) from the bench, then return it tothe normal position. This resets the movementdetection system.

3.1.12 Press [OPEN LID ] and at the same

Iss 1.1

3.3

1CNU03-GB-EFG

3.1.6

3.1.8

3.1.11

time lift the see-through safety cover from thefront.

3.1.13 If power is not avai lable, use theEmergency Release.

3.1.14 To use the Emergency Release,remove the small cap at the left side of the upperhousing and insert a rod such as a pencil orscrewdriver that is at least 100 mm (4in.) long,or longer, into the aperture.

3.1.15 Push the rod inward to release thelock.

3.1.16 Lift the safety cover from the front toopen.

3.1.17 Remove the sponge packing from thetop of the Sealed Head assembly then lift theSealed Head assembly out of the Cytospin®.Remove all the packing pieces.

3.1.18 Pull up the centre button of the lid ofthe Sealed Head until it 'clicks', then lift off thelid.

Note1 The lid of the Sealed Head fits snugly in the

rubber seal of the base. You may need totilt the lid gently before you lift it off.

Iss 1.1

3.4

1CNU03-GB-EFG

3.1.12

3.1.14

3.1.17

3.1.18

4.1.1 To Open the Sealed Head, pull up thecentre button of the Sealed Head lid until it'clicks'. Remove the lid. If necessary, hold thelid of the Sealed Head with one hand and pullup the centre button with the other.

4.1.2 To Close the Sealed Head, place thelid in position on the seal of the base. Makesure that the lid sits on the seal correctly. Pushdown the centre button to secure.

Do not open or close the SealedHead assembly while i t is

installed in the instrument or you maydamage the instrument.

4.1.3 To Load the Cytoclip™ slide clip:

i fit the glass slide (1), then

ii fit the filter card (2), then

iii fit the re-usable sample chamber (3).

iv Pull up the spring (4) and press it intothe two retaining hooks to hold thechamber in place.

Note1 The Cytofunnel® is loaded by pipette with

between 0.1 ml and 0.5 ml (max.) ofcells in suspension.

2 Detailed instructions for loading theCytospin® are given in Appendix 1 -METHODOLOGY GUIDELINES

Iss 1.1

4.1

1CNU04-GB-EFG

4.1.2

4.1.31

2

3

4.1.1

WARNING

4

4.1.4 To Unload the Cytoclip™ hold theslide clip firmly. Push the spring against theslide to release it from the hooks. Move thespring away from the hooks and allow thesample chamber to pop out of the clip.

4.1.5 Disposable Cytofunnels have filtercards permanently attached which simplifiesloading and unloading the Cytoclip™. Whenunloading, discard the Cytofunnel® directly intothe waste receptacle.

4.1.6 To Load the Sealed Head remove thelid and place up to 12 assembled Cytoclips intothe slots. Each Cytoclip™ must be free toswing between the tilt and upright positions.

4.1.7 Make sure that the Cytoclips areevenly distributed so that Cytospin® is notout of balance. Replace the lid and pushdown the centre button to lock.

4.1.8 To Install the Sealed Head inCytospin ® lift the Sealed Head by its centreknob and place it carefully in position on thetapered boss. Do not wipe the silicone greasefrom the tapered boss. Close the see-throughsafety cover.

4.1.9 To Remove the Sealed Head. Allowrotation to stop then press [OPEN LID]. Openthe safety cover and remove the Sealed Headto a biologically safe cabinet. Only open theSealed Head rotor assembly in a biologicallysafe cabinet.

Iss 1.1

4.2

1CNU04-GB-EFG

4.1.4

4.1.5

4.1.8

5.1.1 Control of Cytospin® 3 is by membranetype, touch sensitive, push-buttons distributed onthree panel sections. The push-buttons lightwhen selected and numerical displays showthe current status of the instrument.

5.1.2 The PROGRAM MODE section of thepanel enables the operating parameters forSPEED, TIME and ACCELeration to be set andindicated. Programming is described inSection 6.

5.1.3 The ALARM section of the panel hasindicators that light if a fault occurs.

5.1.4 The CONTROL section of the mainpanel provides means for overall control of theinstrument with OPEN LID, START and STOPfacilities.

Iss 1.1

5.1

1CNU05-GB-EFG

5.1.2

5.1.3

5.1.4

5.1.1

5.2.1 Numeric keys are used in conjunctionwith the function keys, such as SET TIME, toset a desired parameter.

5.2.2 SET SPEED enables an operatingspeed of between 200 to 2000 r.p.m to be set.The selected speed is displayed in the SPEEDdisplay.

5.2.3. SET TIME is the durat ion thatCytospin® operates at the set speed. Therange available is between 1 and 99 minutes.The actual time selected is displayed in theTIME display.

5.2.4 ENTER confirms the key-pressselections.

5.2.5 CANCEL removes the previous key-press entry.

5.2.6 PROGRAM displays a numberbetween one and nine to identify each of thenine individual programs that can be stored.

Iss 1.1

5.2

1CNU05-GB-EFG

5.2.4

5.2.1

5.2.2

5.2.3

5.2.5

5.2.6

5.2.7 LOAD is used to select a program fromthe memory. The speed, time and accelerationsettings of the selected program are displayed.

5.2.8 SAVE transfers program parameters tomemory. The program can subsequently beloaded to view or to use.

5.2.9 SPEED displays the speed selected forthe program being entered.

5.2.10 TIME displays the durat ion of theselected program.

5.2.11 ACCEL selects the rate at which theSealed Head is accelerated up to the setspeed. The options are HIGH, MEDium, orLOW.

5.3.1 The BAL indicator, when lit, shows thatthe Sealed Head assembly is out of balance.

5.3.2 The ‘ ! ‘ alarm indicator lights to showthat Cytospin® has operated over speed andhas stopped for safety.

Iss 1.1

5.3

1CNU05-GB-EFG

5.2.8

5.2.9

5.2.11

5.2.7

5.3.1

5.3.2

5.2.10

5.3.3 A ‘Lid Close’ safety sequence looks forcorrect opening and closing of the safety cover.The LID LOCK indicator lights when

i the instrument is first switched on,ii if the safety cover is not closed correctly.

The instrument does not start when LID LOCKis lit. Press [OPEN LID], open and raise thetop cover at least 25mm (1”), then close the lidfirmly to cancel this alarm.

5.3.4 The BAL and ! alarms flash if Cytospin®

is physically moved from its set position for anyreason. I f Cytospin ® is moved whi le i t isswitched off, the BAL and ! alarms flash whenCytospin® is next switched on. Tilt the instru-ment from the front (lift 15mm - ½”), then lower,to cancel this alarm.

5.4.1 [OPEN LID] releases the interlock ofthe safety cover when the Sealed Head is static.

5.4.2 [START] starts the selected program.

5.4.3 [STOP] discontinues the currentprogram.

5.5.2 The tones vary according to status:

Short Tone = Key press acknowledged.

Long Tone = Invalid key press.

Pulsed tone = Alarm condition.

3 second tone= End of Cycle (ready to open); Cover Open/Close required.

Iss 1.1

5.4

1CNU05-GB-EFG

5.3.3

5.3.4

5.4.1

5.4.2

5.4.3

6.1.1 Proceed as follows:

Iss 1.1

6.1

1CNU06-GB-EFG

NOTE1 If the set time, or set speed, are outside the

limits then the long tone sounder operatesand Cytospin ® 3 does not accept the entrywhen [ENTER] is pressed.

PRESS RESULT

PowerSwitch

[OPEN LID]

[SET TIME]

[Number]

[ENTER]

[SET SPEED]

[Number]

[ENTER]

[HIGH]/[MED]/[LOW]

The displays show ' 0' ;HIGH accel. indicator is lit.Tone sounds.LID OPEN is lit

Open and close lid.

The TIME display flashes ' 0'.

The TIME display flashes thenumber which must be between 1and 99 minutes.

The TIME display shows thenumber.

The SPEED display flashes ' 0'.

The SPEED display flashes thenumber which must be between200 and 2000.

The SPEED displayshows thenumber.

The ACCEL. indicator push-button shows the selection.

6.2.1 When a program is entered press[START] to run the program.

6.3.1 To save an entered program tomemory:

i Press [SAVE].ii Enter a program number (1-9) [ n ].iii Press [ENTER].

6.3.2 Programs can not be entered or savedwhile Cytospin® 3 is running.

6.4.1 To run a program that is stored inmemory:

i Press [LOAD] .ii Enter a program number (1-9) [ n ].iii Press [ENTER].iv Press [START].

6.5.1 To view a program that is held in thememory:

i Press [LOAD] .

ii Enter a program number (1-9) [ n ].

Iss 1.1

6.2

1CNU06-GB-EFG

6.2.1

6.3.1

6.4.1

6.5.1

6.6.1 Install the loaded Sealed Head in theCytospin® and press [START]. It may beconvenient to load a second Sealed Head in abiologically safe area while the first is in use.

6.6.2 Swap the Sealed Head units at the endof a completed cycle. Press [START] to repeatthe program.

6.7.1 Press [CANCEL] to cancel the lastkeystroke entry.

6.8.1 Press [STOP] to terminate a programbefore its set time. The STOP indicator lightsand the SPEED display shows the SealedHead losing speed.

6.8.2 When the speed of the Sealed Headdrops below 50 r.p.m. the SPEED display flash-es '50' until the Sealed Head is stationary. The'End of Cycle' sounder then operates.

6.8.3 The safety cover remains locked andall pushbuttons of the control panel are dis-abled until the Sealed Head is stationary.

6.8.4 The control panel is re-enabled afterthe 'End of Cycle' tone.

Iss 1.1

6.3

1CNU06-GB-EFG

6.6.2

6.7.1

6.8.1

6.8.2

Iss 1.1

6.4

1CNU06-GB-EFG

7.1.1 Cytospin® 3 uses centrifugal force todeposit a monolayer of cells in a defined areaon glass slides. It effectively by-passes thedifficulties normally associated with depositionsobtained by direct smear or filtration.

7.1.2 Maximum protection for the operator isensured by completely containing potentiallyhazardous specimens in a Sealed Headassembly.

7.1.3 Good laboratory practice requires theuse of a biological safety cabinet for loadingand unloading the Sealed Head. The SealedHead should be returned to the cabinet afterspinning, before it is opened.

The use of a biologically safecabinet when loading or

un-loading the Sealed Head is particularlyimportant if the samples underinvestigation contain, or could contain,pathogenic micro-organisms.

7.1.4 Pencil marking of frosted end slides isa common method for identifying samples andis a recommended practice that takes intoaccount the expected use of the slide.

7.1.5 The instrument should be regularlycleaned, disinfected and sterilized as describedin Section 8.

Iss 1.1

7.1

1CNU07-GB-EFG

7.1.2

7.1.4

WARNING

7.1.6 Each laboratory has its own techniquesfor preparing cells. The accompanying Table -CYTOSPIN 'g' FORCES - provides helpfulinformation regarding the forces generated inCytospin® 3. The 'g' forces quoted apply at theface of the slide that receives the cells.

7.2.1 The paper:

Use of the Cytospin for Hematology andother Clinical Microscopy Specimens,

prepared for Shandon Scientif ic by NickySherwood, MT (ASCP) and Joanne Cornbleet(MD) of the Clinical Hematology Laboratory,Stanford University Medical Center, is includedin English as Appendix 1 at the back of thismanual.

7.3.1 If the BAL alarm lights while Cytospin®

is operating,

i the program terminates and the pulsedtone alarm sounds (the figure in thePROGRAM display changes to a diag-nostic code and should be ignored).

ii the alarm sounds until rotation ceases,and the Sealed Head is stationary.

iii use the power ON/OFF switch to switchthe instrument OFF then ON again.

iv use [OPEN LID] to release the safetycover. Remove the Sealed Head to abiologically safe cabinet before openingit . Check that the load is correct lybalanced and evenly distributed.

Iss 1.1

7.2

1CNU07-GB-EFG

SPEED FORCE(r.p.m.) (g)

100150200250300350400450500550600650700750800850900950

100010501100115012001250130013501400145015001550160016501700175018001850190019502000

1.132.544.527.06

10.1613.8318.0622.8628.2334.1540.6547.7055.3263.5172.2681.5791.45

101.90112.90124.48136.61149.32162.58176.41190.81205.77221.29237.38254.03271.25289.03307.38326.29345.77365.81386.41407.58429.32451.62

v Close the Sealed Head assembly beforeyou remove it from the biologically safecabinet. Load the Sealed Head unit inthe Cytospin ® and run the programagain.

7.3.2 If BAL and ! both flash, lift the front ofthe instrument approximately 15mm (½ ins)from the bench then return it to its set position.(see 5.3.4)

7.3.3 If the ! alarm indicator lights while theCytospin is operating:

i the program terminates and the pulsedtone alarm sounds (the figure in thePROGRAM display changes to adiagnostic code and should be ignored).

ii the alarm sounds until the Sealed Headis stationary.

iii use the power ON/OFF switch to switchthe instrument OFF then ON again.

iv use [OPEN LID] to release the safetycover. Check that the Sealed Headassembly is installed correctly. If thealarm persists contact your ThermoShandon Service department.

Iss 1.1

7.3

1CNU07-GB-EFG

7.3.2

7.3.4 If LID LOCK is lit

i press [OPEN LID] and open the safetycover at least 25mm (approximately1in.). Close the safety cover and tryagain.

Note1 Cytospin® is normally kept switched on and

ready for use. Each time the Sealed headis installed, the safety system detects theopening and closing of the safety cover inthe correct sequence and allows Cytospin®

to start when [START] is pressed.

2 If the Sealed Head is instal led beforeCytospin® is switched on, the safety systemcannot detect the correct safety coveropening and closing sequence so [START]is disabled and the LID LOCK alarm lights.

Iss 1.1

7.4

1CNU07-GB-EFG

8.1.1 Cytospin ® is designed for easymaintenance and most fixed components suchas the Safety Cover, the Bowl Liner, the ControlPanel and the metal housing are cleaned usinga proprietary mild detergent solution appliedwith a soft cloth.

8.1.2 All components and accessoriesthat are likely to become contaminated are alsoeasily cleaned with proprietary mild detergentsolutions after sterilization. It is recommendedthat the suggested methods for sterilization areused.

8.2.1 Make sure that the active ingredi-ents of any cleaning materials or disinfectantsthat you propose to use are compatible with thematerials used in the construction of Cytospin®

and its accessories. If you are not sure, pleasecheck with Thermo Shandon servicedepartment first.

8.2.2 Most proprietary disinfectants incommon laboratory use, such as Clorox®, orcommercial disinfectants diluted with 0.3%bicarbonate buffer at 7.0 to 8.0 pH, should besuitable.

8.2.3 Al low dis infectant to contact acontaminated surface for at least one hour,where practicable, to ensure sterilization.Twenty hours exposure is required todestroy spores of Bacillus subtilus 1.

Iss 1.1

8.1

1CNU08-GB-EFG

WARNING

WARNING

WARNING

WARNING

WARNING

WARNING

Always wear protective gloves whenyou clean or sterilize Cytospin ® toprotect yourself against infection orthe effects of chemicals.

Do not clean or sterilize by methodsthat are not recommended byThermo Shandon.

Do not use any chemicals that interactwith materials of manufacture. If indoubt, check with Thermo ShandonService department.

Remove the mains plug from thesupply socket before you clean thefixed components of the instrument.

Phenol and Hypochlorites in strongsolution will damage the instrumentand its accessories.

Do not use abrasive compounds ormetal components to cleanCytospin ® or its components andaccessories.

8.3.1 FRONT PANEL

Note1 Disconnect Cytospin ® from the mains

supply.

FREQUENCYWeekly or after spillage

CLEAN: Use warm soapy water on dampenedcloth or sponge.

Apply 10% commercial bleach inwater on dampened cloth or sponge.

Finish with liquid spray or furniturepolish.

AVOID Abrasive powders.Xylene, toluene or similar solvents.

8.3.2 SAFETY COVER


supply.

FREQUENCYDaily or after serious spillage



Finish with liquid spray or furniturepolish.


Iss 1.1

8.2

1CNU08-GB-EFG

8.3.1

8.3.2

8.3.3 BOWL LINER


supply.

FREQUENCYWeekly or after spillage




8.3.4 SEALED HEAD ASSEMBLY

FREQUENCYDaily and immediately after any

serious spillage.

STERILIZEAutoclave all the Sealed Head, itscomponents and accessor ies at121°C (250°F) for 15 minutes.Unlock the lid so that steam can fullypenetrate the interior.

Alternatively, submerge the SealedHead and i ts components andaccessories in a 10% solut ion ofcommercial bleach in water for notless than one hour.

CLEAN: After sterilization wash the SealedHead and i ts components andaccessories in warm soapy water. Dryin an oven at a temperature notexceeding 65°C (149°F).

Iss 1.1

8.3

1CNU08-GB-EFG

8.3.3

8.3.4

8.3.5 SEALED HEAD BASE


serious spillage.

STERILIZEAutoclave at 121°C (250°F) for 15minutes.

Alternatively, submerge in a 10%solut ion of commercial bleach inwater for not less than one hour.

CLEAN: After sterilization wash the SealedHead base in warm soapy water.Rinse in clear water, then dry.

AVOID the use of hard brushes.

8.3.6 SLIDE CLIP SUPPORT PLATE

Note1 Undo and remove the two thumb screws

then lift up and remove the Support Plateto gain access to its underside.

FREQUENCYDaily.


Alternatively, submerge in a 10%solut ion of commercial bleach inwater for not less than one hour.

CLEAN: After sterilization wash the Slide ClipSupport Plate in warm soapy water.Rinse in clear water, then dry.

AVOID the use of hard brushes.

Iss 1.1

8.4

1CNU08-GB-EFG

8.3.6

8.3.5

8.3.7 SEALED HEAD LID


serious spillage.

Weekly Remove the silicone rubberseal from around the rim ofthe lid and clean the surfaceof the lid. Replace the seal.

Monthly grease the locking bal lbearing assembly.

Annually fit a replacement seal.


Alternatively, submerge in a 10%solution of commercial bleach inwater for not less than one hour.

CLEAN: After sterilization wash the SealedHead lid in warm soapy water. Rinsein clear water, then dry.

AVOID the use of hard brushes.abrasive powders.Xylene, toluene or similar solvents.

Iss 1.1

8.5

1CNU08-GB-EFG

8.3.7

8.3.8 CYTOCLIP™ SLIDE CLIP

Note1 For Slide Clip, Part No 5991052M, snap

open the spring: for spring loaded SlideClip, Part No 5991017M, turn thepressure release screw fully counterclockwise to release the spring pressure.

FREQUENCYDaily



CLEAN: Rinse in clear water and hot air drybefore use.

8.3.9 REUSABLE SAMPLE CHAMBER

FREQUENCYAfter use.



CLEAN: Use a cotton tipped applicator swaband hot soapy water to clean theoutlet port and funnel. Rinse in cleanwater then dry.

Iss 1.1

8.6

1CNU08-GB-EFG

8.3.8

8.3.9

AVOID Immersion in disinfectant for morethan 5 minutes.

Phenolic disinfectants.

Autoclaving the plastic cap.

The use of a wire brush.

Note1 Discolorat ion due to repeated

autoclaving does not adversely affect theperformance of the Sample Chamber.

8.3.10 FILTER CARDS

Note1 The Filter Cards are disposable, single -

use i tems that are designed to bediscarded immediately after use.However, they must be rendered safe bysterilization before being consigned fordisposal.

FREQUENCYAfter use.

STERILIZEDiscard into a phenolic disinfectant,or,

Autoclave at 121°C (250°F) for 15minutes.


DISPOSEin the approved manner according tolocal procedures.

Iss 1.1

8.7

1CNU08-GB-EFG

8.3.10

8.4.1 Three flexible seals are fitted in theSealed Head - the Cone Seal (1), an 'O' RingSeal (2), and a Lid Seal (3).

8.4.2 All the seals are designed to withstandnormal cleaning and sterilization as part of theroutine maintenance of the Sealed Head.However, the seals eventually become worn,stretched, or degraded by the act ion ofchemicals over a period of time and should bereplaced annually.

8.4.3 To Remove the Sealed Head ConeSeal (1), simply pull the old seal offthe cone.

8.4.4 To Fit the Sealed Head Cone Seal(1). Stretch the replacement ConeSeal (Part No P07431), with the thinlip uppermost, onto the collar of thecentre cone.

8.4.5 To Remove the Sealed Head 'O'Ring Seal (2), Use a screwdriver toundo the four screws that secure thecone to the base, then remove thecone and l i f t the 'O' r ing from itsgroove.

Iss 1.1

8.8

1CNU08-GB-EFG

1 2 3

8.4.1

8.4.3

8.4.5

8.4.6 To Fit the Sealed Head 'O' RingSeal (2). Fit a replacement 'O' RingSeal (Part No P07525) in its groove inthe cone. Make sure that the 'O' ringfits correctly in its groove then fit thecone to the base. Fit, then tighten, thefour screws.

8.4.7 To Remove the Sealed Head LidSeal (3), pull the old seal off the rimof the bowl.

8.4.8 To Fit the Sealed Head Lid Seal (3).Place the replacement Lid Seal (PartNo P07516) on the rim of the bowlthen push the lip of the seal over thevertical edge at the periphery of thebowl. Make sure that the seal fitsuniformally around the rim.

Iss 1.1

8.9

1CNU08-GB-EFG

8.4.7

8.4.8

Iss 1.1

8.10

1CNU08-GB-EFG

Displays not lit on control panel

9.1.1 Correct service and maintenance is essential for the long term serviceability of precision engineeredproducts such as Cytospin® 3. We strongly recommend that a Thermo Shandon Service Contract is used toensure future reliability, and consistency of performance.

9.1.2 Table 1 shows remedial action to be taken if Cytospin ® fails to operate.

9.1.3 Tables 2 to 5 relate to processing problems and suggested solutions with respect to the preparation ofcells by cytocentrifugation.

Iss 1.1

9.1

1CNU09-GB-EFG

SYMPTOM CAUSE REMEDY

Programs do not run.

BAL alarm lit.

alarm lit.

BAL and alarms flash together.

1 No Power Supply.2 Mains fuse blown.3 Internal Instrument

Fuse blown.

1 Stopped at instrument.2 Incorrect programming.

1 Load out of balance.

1 Overspeed.

1 Cytospin travelling.

1 Check the Mains supply.2 Replace the mains fuse.3 Replace the instrument fuse (Note only a

technically competent person shouldreplace fuses).

1 Check load distribution. Reset alarm.(see 7.3.2).

TABLE 1 - EQUIPMENT STATUS CHECKS

1 Switch OFF then ON again.2 Check sample carrier distribution.3 Check Sealed Head for damage.

1 Press [START].2 Check ranges 200 -2000 r.p.m.

1 - 99 minutes.

1 Switch OFF then ON again (see 7.3.3).2 Check that the Sealed Head is installed

correctly.3 Contact Thermo Shandon Service i f

problem persists.

LID LOCK lit 1 Lid sequence interlock. 1 Open and close lid. (see 7.3.4).

Iss 1.1

9.2

1CNU09-GB-EFG

TABLE 2 QUALITATIVELY ABERRANT CYTOCENTRIFUGATION RESULTS

Poorly preservedcells.

1 Preserved poorly in vivo.2 Lengthy delays between collection and

preparation.3 Cells suspended in normal saline.

1 Request repeat specimen.2 Minimize delays (e.g. less than 4 hr.

Refrigerate if longer).3 Use balanced electrolyte solution.

PROBLEM CAUSE SOLUTION

Cells small indiameter; opticallydense.

1 Cells collected in high proportion ofstrong alcohol.

2 Alcohol added to sample chamberbegins to rise through the cell suspen-sion and causes cell shrinkage when itmixes.

1 Collect unfixed fresh specimens, or mixwith equal volume of 50% ethanol.

2 Add less alcohol; add alcohol carefully.

RBCs hemolysed;ghosts remain.

1 Alcohol was mixed with cell suspension. 1 Add alcohol carefully to sample chamber.

BEFORE CYTOCENTRIFUGATION

DURING CYTOCENTRIFUGATION

Cells air dried. 1 Cell suspension medium absorbedcompletely by filter card.

1 Fill cylindrical sample chamber beforecytocentr i fugat ion; use Cytospin®

Col lec t ion F lu id ; reduce thecytocentrifugation time and / or speed.

Cel ls a i r dr iedaround peripheryof collection area.

1 Cell suspension medium almostcompletely absorbed by filter card.

1 Increase specimen volume up to 0.5ml;use Cytospin® Collection Fluid; reducecytocentrifugation time and/or speed.

Cells air dried. 1 Film of liquid over the cells allowed toevaporate dur ing br ief intervalbetween unloading and immersion inalcohol.

1 Move quickly to avoid evaporation ofprotective film from over the cells.

Cel ls air dr iedaround peripheryof collection area.

1 Film of liquid is thinnest at its edgesand evaporates before the thick cen-tral area.

1 Immerse cells in alcohol before air dry-ing progresses on to periphery of cellarea.

AFTER CYTOCENTRIFUGATION

Disrupted air driedpale cells, resem-ble 'basket cells' ofhematology.

1 Fragile cells air dried and exaggeratedby centrifugal force.

1 Request repeat specimen. Do not allowto air dry.

Iss 1.1

9.3

1CNU09-GB-EFG

TABLE 3 QUANTITAVELY ABERRANT CYTOCENTRIFUGATION RESULTSNumber of Cells

No cells 1 Exit port blocked by filter card. 1 Seat filter card to foot of side clip.


Abnormal cells inspecimen but noton cytocentrifugedpreparations.

1 Abnormal cells, usually larger andheavier than normal cells, sedimentto bottom of concentrate. May bemissed if not resuspended completelyfollowing conventional centrifugation.

1 Apply centrifuge tube with cell concentrateand several ml of balanced electrolytesolution to vortex mixer and completelyresuspend cells.

Too few cells. 1 Too few cells in raw specimen.


2 Too few cells added to samplechamber.

2 Base sample size on cell count of drop ofresuspended cell concentrate.

3 Sparsely populated specimen mayhave filled the cylindrical and conicalportions of the sample chamber.

3 Enrich specimen as described above.

4 Partially filled cone raised level ofspecimen in cylinder to filter cardlevel where cells can be absorbed.

4 Do not allow distal boundary of specimen totouch filter card before cytocentrifugation.

5 Cells crowded out by precipitatedhyaluronic acid in joint fluid.

5 Dissolve hyaluronic acid precipitate withpinch of hyaluronidase.

1 Enrich by conventional centrifugation.Resuspend cel ls in 1-2ml balancedelectrolyte solution. Combine contents ofmultiple centrifuge tubes of same specimenwhen possible. microscopically examinedrop of resuspended cell concentrate;Base sample size on cell count. Requestrepeat specimen; suggest ways toincrease cellular harvest.

6 Cells crowded out by precipitatedphosphate salts in urine.

7 Cells crowded out by erythrocytes. 7 Saponize specimen.

6 Mix several drops of glacial acetic acid tolower the pH and redissolve the alkaline pHdependant preciptated phosphate salts.

8 Slide is between exit port and filtercard .

8 Load in correct sequence; samplechamber filter card side.

Too many cells. 1 Too much densely populated cellsuspension added to sample chamber.

1 Microscopical ly examine drop ofresuspended cell concentrate; dilute upto 10x if necessary; base sample size oncell count or derive it from hematologicalcounting chamber.

2 Do not rely on visual estimates of specimenappearance.

Iss 1.1

9.4

1CNU09-GB-EFG

TABLE 3 QUANTITAVELY ABERRANT CYTOCENTRIFUGATION RESULTSNumber of Cells (Continuation)

Too few cells. 1 Exit port blocked by misaligned filtercard.

1 Check alignment as seen through windowfrom back of slide clip; seat filter card.


2 Cells lost through gap between exitport and filter card.


1 Suspension medium absorbedincompletely as a result of filter cardbecoming clogged by debris inpreviously non-centrifuged specimenand/or pores collapse from too muchpressure from spring or excessivecentrifugal force

1 Centrifuge specimen at 3000 r.p.m. for 10min. to sediment cells and leave debris insuspension to be discarded wi thsupernatant. Do not wet blotter beforecytocentr i fugat ion. Cytocentr i fugespecimen at 1000 r.p.m. for 6-10 min.

1 Unlikely; though check assembled unit foralignment.

AFTER CYTOCENTRIFUGATIONToo few cells.

2 Unabsorbed suspension medium caninduce cell wash-off.

2 Unload horizontal sample chamber, cellside up. Allow blotter to absorbexcessliquid. Lift chamber and blotter awayfrom slide. Lay slide flat until a thin filmremains; immerse in fixative.

TABLE 4 UNUSUAL PATTERN OF CELL POPULATION DISTRIBUTION

Crescent shapeddistribution.

1 Cylinder filled incompletely. 1 Fill cylinder completely.


2 Cells settle in cylinder if prolongeddelay before cytocentrifugation.


2 Rapidly load sample chambers and begincytocentrifugation immediately.

Display areadisplaced fromlabel end.

1 Slide not seated to foot of slide clip. 1 Seat slide to foot of slide clip.

1 Slide placed in slide clip label enddown.

1 Insert slide label end up.Display areadisplaced towardslabel end.

1 Slides loaded backwards. 1 Orient slide with label facing exit port.Cells on undersideof slide

Iss 1.1

9.5

1CNU09-GB-EFG

1 Thinly layered cells too wet and areeither pushed up the sl ide uponimmersion in alcohol, or slide downthe slide following immersion.

1 Let the suspension medium evaporatealmost completely.


Cell populat ionstreams towardslabel end or toopposite end.

TABLE 4 UNUSUAL PATTERN OF CELL POPULATION DISTRIBUTION(Continuation)


1 Thickly layered cells, margins nearlydry but centre remains very wet andpromotes cell wash-off.

1 Add less cell suspension.Circular band ofcel ls, acel lularcentre, 'Bull's Eye'distribution.

TABLE 5 SUMMARY To predictably produce cytocentrifuged preparations that exhibit within a 32mm² circle a representative sampleof randomly distributed, uncrowded, monolayered, flattened cells that are well preserved and displayed, thematerials and methods that follow are recommended:


1 Use Unfixed fresh cell suspension.2 Saponinize bloody cell suspensions.3 Equalize differences in cell suspensions4 Control the number of cells.5 Use clean Micro slides.6 Use balanced electrolyte solution.7 Keep cell suspension from the filter card.8 Fill sample chambers with similar volumes.

1 Use cell suspensions collected in alcohol.2 Use bloody cell suspensions.3 Cytospin un-processed cell suspensions.4 Estimate number of cells.5 Use frosted or albumenized micro slides.6 Use normal saline.7 Let the cell suspension touch the filter card.8 Use significantly different volumes.


1 Centrifuge specimen at 1000 r.p.m.2 Centrifuge for 3 - 4 minutes until nearly dry.

1 Centrifuge too rapidly.2 Centrifuge too briefly, or for too long.


1 Keep the cells slightly wet.2 Immerse immediately the cytological

preparation in 95% ethanol for ethanolpreps.

3 Allow the cellular monolayer to air-drybefore staining when using Cytospin ®

Collection Fluid

1 Allow the cells to air dry unless intended.2 Let too much liquid remain on the cells.

3 Immerse in 95% ethanol until the cell monolayersprepared with Cytospin Collection Fluid have driedsufficiently - unless otherwise specified.

Iss 1.1

9.6

1CNU09-GB-EFG

10.1.1 Physical Height 215mm (8 1/2 in)Height - Clearance 560mm (22 in)Width 385mm (15 1/4 in)Width - Clearance 405mm (16 in)Depth 495mm (19.5 in)Depth - Clearance 556mm (21.7/8 in)Weight (Kg) 18 Kg (39 3/4 lb)Mounting Bench - 4 fixed feet

10.1.2 Electrical Power 150 VAVoltages - (factory set) 110 - 120 V ~

220 - 240 V ~Frequency 50/60 HzFuse Type:

Mains Plug 5 Amp (UK instruments only)110 - 120 V T2.0 Amp Slow Blow220 - 240 V T0.8 Amp Slow Blow

10.1.3 Switch Convention l = Power OnO = Power Off

10.1.4 Indicator Panel All Models Membrane type, touch sensitive switches and LED displays.

10.1.5 Capacities All Models Sample Chamber 0.1 - 0.5 mlMegafunnel 6.0 ml.

Note1 This instrument is intended for use with body fluids. It is not necessary to de-rate the instrument for different

densities of sample provided that the instrument is used for its intended purpose.

10.1.6 EnvironmentGeneral Indoor use only.Temperature +5°C to + 40°CHumidity 80% max for temperatures < 31°C

50% max for temperatures 31°C to 40°C(Non condensing environment)

Altitude Up to 2000mPollution Degree 2 (as defined in IEC1010)Installation Category II (as defined in IEC1010)

10.1.7 Operating Speed Range 200 - 2000 r.p.m.

10.1.8 Safety Interlocks Safety Cover sequence interlock.Safety Cover locked closed while centrifuging.Controls inoperable while the rotor is turning.Drive motor cut off if instrument moves.

10.1.9 Noise The sound pressure level at workstations does not exceed 70 dB.

Iss 1.1

10.1

1CNU10-GB-EFG

DESCRIPTION CATALOGUE NUMBER

International US

Iss 1.1

10.2

1CNU10-GB-EFG

CYTOSPIN® Unit - each

Packing and Carton - each

Sealed Head (Slide Clips not supplied) - each

Stainless steel Cytoclip™ Slide Clip - 6 pack

Stainless steel Cytoclip™ Slide Clip - 2 x 6 pack

Re-usable (Autoclavable) Sample Chamber - each

Operator Guide handbook - each

Silicone Rubber Head Seals- - 1 x 3 pack

Cytofunnel® disposable double white Sample Chambers with caps - box 25

Cytofunnel® disposable single white Sample Chambers with caps - box 50

Cytoslide™ Regular Microscope slides, single circle, frosted end - box 100

Cytoslide™, double circle, Sample - 2 x box 25

Consumables Catalog - each

Sample pack containing6 x Collection Fluid (60ml cup)1 x Collection Fluid (500ml bottle)1 x Cell Fixx™ Spray (50ml)

Warranty Card - each

Cytospin® Wall Chart

Extended Service Contract Offer - each

Delivery Questionnaire - each

74000001

59910018

59910052

5991002

74010121

59910019

59910039

59910040

59910051

7400002

1612084

5991018

5991052

7401121

5991019

5991039

5991040

5991051

5991058

31202

1950425

1000291

1001436

1001437

Thick White filter cards for volumes of 0.5 ml - box 200

Brown filter cards - box 200

TPX Sample Chamber - each

Closure Caps - 12 pack

Cytofunnel® disposable single white Sample Chambers with caps - box 50

Cytofunnel® single Sample Chambers (white, bulk pack) - box 500

Cytofunnel® disposable double white Sample Chambers with caps - box 25

Cytofunnel® single Sample Chambers (brown) - box 50

Cytofunnel® double Sample Chambers (white, bulk pack) - box 500

Cytofunnel® single Sample Chambers, Sterile - box 25

Cytofunnel® double Sample Chambers, Sterile - box 50

Megafunnel large volume Sample Chamber (6.0 ml capacity) - box 25

Cytoslide™ Regular Microscope slides, single circle, frosted end - box 100

Cytoslide™ Coated Microscope slides, single circle, frosted end - box 100

Cytoslide™ Regular Microscope slides, double circle, frosted end - box 100

Cytoslide™ Coated Microscope slides, double circle, frosted end - box 100

Cytoslide™ Black Mask Microscope slide for immunofluorescence - box 100

Regular Microscope slides, Plain, (sold only by gross)(USA) - box 72

Regular Microscope slides, Frosted End, (sold only by gross)(USA) - box 72

Regular Microscope slides, Superfrost®, (sold only by gross)(USA) - box 72

59910022

59910023

59910021

59910025

59910040

1102548

59910039

59910043

1102547

5991044

5991042

5991028

59910051

59910056

59910054

59910055

59910057

5991022

5991025

5991040

1102548

5991039

1102547

5991044

5991042

5991028

5991051

5991056

5991054

5991055

5991057

1450

146012

147019

Iss 1.1

10.3

1CNU10-GB-EFG


International US

Iss 1.1

10.4

1CNU10-GB-EFG

Cytospin ® Collection Fluid in 4 litre container with pump - each

Cytospin ® Collection Fluid in 120 ml cups 1/2 filled - case 50

Cytospin ® Collection Fluid in 500 ml bottles (USA) - 2 pack

Cytospin ® Collection Fluid in 10 litre bottles - 2 pack

Cell-Fixx™ spray fixative in 50 ml bottle(USA) - each

Cell-Fixx™ spray fixative in 50 ml bottles - 6 pack

Mucolexx Transport Medium Fluid in 500ml bottles - 4 pack

Cytoblock™ Cell Block Preparation System - each

9990301

9990321

9990315

9990310

9990325

9990326

9990370

7401050

67680001

67680017

67680016

99900326

9990370

74010150


International US

We are proud of our quality and reliability, and of our after-sales service. We continuously strive toimprove our service to our customers.

Please ask your distributor or representative about Service Contracts which can keep your purchase inpeak condition for many years to come.

Warranty provisions necessarily vary to comply with differences in national and regional legislation, andyou can find details in your delivery documents or from your dealer or representative.

Iss 1.1

11.1

1CNU11-GB-EFG

Iss 1.1

11.2

1CNU11-GB-EFG

This product conforms with the essential requirements of the following directives;

EMC Directive 89/336/EEC ( as amended by 92/31/EEC & 93/68/EEC ).

Low Voltage Directive 73/23/EEC ( as amended by 93/68/EEC ).

This product complies with the following International Standards:

Manufacturer's Name:

Manufacturer's Address:

Product Description:

Product Designation:

Year of Marking (CE):

Declaration of Conformity

Thermo Shandon Limited,

93 - 96 Chadwick Road, Runcorn, Cheshire, WA7 1PRENGLAND


Cytospin® 31996

Issued by: Mr R.Russell-SmithDirector of QualityThermo Shandon Limited

..................................................................................

EMC:Safety:

EN55022 class B

EN50082 - 1

IEC1010-2-020

CAN/CSA Standard C22.2: 1010-1-92

UL Standard: 3101-1

Iss 1.1

12.1

1CNU12-GB-EFG

TOPIC SECTION PAGE

Acceleration . . .To Set . . . . . . . . . . . . . . . . . . . . . . . .5.2.11 . . . . . . . . . . . . . . . . . .5.3Accessories (Sold Separately) . . . . . . . . . . . . . . . . . .10.2 . . . . . . . . . . . . . . . . . . .10.2Alarms. . . . . . . .5.3.. . .5.3

Audible Tones . . . . . . . . . . . . . . . . . .5.5 . . . . . . . . . . . . . . . . . . . .5.4BAL . . . . . . . . . . . . . . . . . . . . . . . . .5.3.1 . . . . . . . . . . . . . . . . . .5.3

' ! ' . . .. . . . . . . . . . . . . . . . . . . . . . .5.3.2 . . . . . . . . . . . . . . . . . .5.3Lid Lock. . . . . . . . . . . . . . . . . . . . . . .5.3.3 . . . . . . . . . . . . . . . . . .5.4BAL and ! . . . . . . . . . . . . . . . . . . . . .5.3.4 . . . . . . . . . . . . . . . . . .5.4

BAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.3.1 . . . . . . . . . . . . . . . . . .5.3

BAL and ! . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.3.4 . . . . . . . . . . . . . . . . . .5.4Bio Hazard Clearance Certtificate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .Appendix 2

Cancel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.2.5 . . . . . . . . . . . . . . . . . . .5.3Certificate of Conformity . . . . . . . . . . . . . . . . . . . . . . .11.2 . . . . . . . . . . . . . . . . . . .11.2Construction . . .2.2 . . .2.2Consumables (Sold Separately) . . . . . . . . . . . . . . . . .10.2 . . . . . . . . . . . . . . . . . . .10.2

CLEANING AND MAINTENANCE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8.1Routine Maintenance . . . . . . . . . . . .8.3 . . . . . . . . . . . . . . . . . . . .8.2

Bowl Liner . . . . . . . . . . . . . . .8.3.3 . . . . . . . . . . . . . . . . . . .8.3Cytoclip™ Slide Clip. . . . . . .8.3.8 . . . . . . . . . . . . . . . . . . .8.6Front Panel . . . . . . . . . . . . . .8.3.1. . . . . . . . . . . . . . . . . . .8.2Reusable Sample Chamber .8.3.9 . . . . . . . . . . . . . . . . . . .8.6Safety Cover . . . . . . . . . . . . .8.3.2 . . . . . . . . . . . . . . . . . . .8.2Sealed Head . . . . . . . . . . . . .8.3.4 . . . . . . . . . . . . . . . . . .8.2Sealed Head - Base . . . . . . .8.3.5 . . . . . . . . . . . . . . . . . . .8.4Sealed Head - Lid . . . . . . . . .8.3.7 . . . . . . . . . . . . . . . . . . .8.5Slide Clip Support Plate . . . .8.3.6 . . . . . . . . . . . . . . . . . . .8.4

CONTENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2

CONTROL PANEL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.1AUDIBLE TONES . . . . . . . . . . . . . . .5.5 . . . . . . . . . . . . . . . . . . . .5.4GENERAL . . . . . . . . . . . . . . . . . . . .5.1 . . . . . . . . . . . . . . . . . . . .5.1PROGRAM MODE SECTION . . . . .5.2 . . . . . . . . . . . . . . . . . . . .5.2CONTROL SECTION . . . . . . . . . . . .5.4 . . . . . . . . . . . . . . . . . . . .5.4ALARM SECTION . . . . . . . . . . . . . .5.3 . . . . . . . . . . . . . . . . . . . .5.3

Continued

Iss 1.1

12.2

1CNU12-GB-EFG

Continuation

TOPIC SECTION PAGE

Cone Seal, To Fit the Sealed Head ~ . . . . . . . . .8.4.4 . . . . . . . . . . . . . . . . . . .8.8To Remove the Sealed Head ~ . . . . .8.4.3 . . . . . . . . . . . . . . . . . . .8.8

Lid Seal, To Fit the Sealed Head ~. . . . . . . . . .8.4.8 . . . . . . . . . . . . . . . . . .8.9To Remove the Sealed Head ~ . . . . .8.4.7 . . . . . . . . . . . . . . . . . . .8.9

'O' Ring Seal To Fit the Sealed Head ~. . . . . . . . . .8.4.6 . . . . . . . . . . . . . . . . . . .8.9To Remove the Sealed Head ~ . . . . .8.4.5 . . . . . . . . . . . . . . . . . .8.8

Cytoclip To Load the ~ . . . . . . . . . . . . . . . . . .4.1.3 . . . . . . . . . . . . . . . . . . .4.1To Unload the . . . . . . . . . . . . . . . . . .4.1.4.. . . . . . . . . . . . . . . . . . .4.2

Cytofunnels Disposable . . . . . . . . . . . . . . . . . . . .4.1.5 . . . . . . . . . . . . . . . . . . .4.2Cytospin Collection Fluid . . . . . . . . . . . . . . . . . . . . . . .10.2. . . . . . . . . . . . . . . . . . . .10.2

DESCRIPTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2.1Operating Principles . . . . . . . . . . . .2.1 . . . . . . . . . . . . . . . . . . . .2.1Construction . . . . . . . . . . . . . . . . . .2.2 . . . . . . . . . . . . . . . . . . . .2.2

Disinfect . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8.1.1 (ii) . . . . . . . . . . . . . . . .8.1Disposable Cytofunnels . . . . . . . . . . . . . . . . . . . . . . .4.1.5 . . . . . . . . . . . . . . . . . . .4.2Duration To Set . . . . . . . . . . . . . . . . . . . . . . . .5.2.3 . . . . . . . . . . . . . . . . . . .5.2

Emergency Release . . . . . . . . . . . . . . . . . . . . . . . . . .3.1.14 . . . . . . . . . . . . . . . . . .3.4End of Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.5.2 . . . . . . . . . . . . . . . . . . .5.4Equipment Status Checks . . . . . . . . . . . . . . . . . . . . . .Table 1 . . . . . . . . . . . . . . . . .9.1

Filter Cards . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8.3.10 . . . . . . . . . . . . . . . . . .8.7Front Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.1 . . . . . . . . . . . . . . . . . . . .5.1

'g' Forces . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7.1.6 . . . . . . . . . . . . . . . . . .7.2

INDEX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12.1

INSTALLATION INSTRUCTIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3.1

Procedure . . . . . . . . . . . . . . . . . . . .3.1 . . . . . . . . . . . . . . . . . . . .3.1

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1.1 . . . . . . . . . . . . . . . . . . . .1.1

Continued

Iss 1.1

12.3

1CNU12-GB-EFG

Continuation

TOPIC SECTION PAGE

LID LOCK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.3.3 . . . . . . . . . . . . . . . . . . .5.3.4

Loading . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . listed under subject.

MAINTENANCE, CLEANING AND ~ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8.1General . . . . . . . . . . . . . . . . . . . . . . .8.1 . . . . . . . . . . . . . . . . . . . .8.1Recommended Materials . . . . . . . . .8.2 . . . . . . . . . . . . . . . . . . . .8.2Routine Maintenance . . . . . . . . . . . .8.2 . . . . . . . . . . . . . . . . . . . .8.1

Bowl Liner . . . . . . . . . . . . . .8.3.3 . . . . . . . . . . . . . . . . . . .8.3Cytoclip™ Slide Clip . . . . . .8.3.8 . . . . . . . . . . . . . . . . . . .8.6Front Panel . . . . . . . . . . . . .8.3.1 . . . . . . . . . . . . . . . . . . .8.2Reusable Sample Chamber .8.3.9 . . . . . . . . . . . . . . . . . .8.6Safety Cover . . . . . . . . . . . .8.3.2 . . . . . . . . . . . . . . . . . . .8.2Sealed Head . . . . . . . . . . . .8.3.4 . . . . . . . . . . . . . . . . . .8.2Sealed Head - Base . . . . . .8.3.5 . . . . . . . . . . . . . . . . . . .8.4Sealed Head - Lid . . . . . . . .8.3.7 . . . . . . . . . . . . . . . . . . .8.5Slide Clip Support Plate . . .8.3.6 . . . . . . . . . . . . . . . . . . .8.4

Methodology Guidelines Appendix 1

Open Lid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.4.1 . . . . . . . . . . . . . . . . . . .5.4

OPERATING . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7.1Introduction .7.1 . . . . . . . . . . . . . . . .7.1Methodology Guidelines . . . . . . . . .7.2 . . . . . . . . . . . . . . . . . . . .7.2To Reset Cytospin ® . . . . . . . . . . . . .7.3 . . . . . . . . . . . . . . . . . . . .7.2

BAL . . . . . . . . . . . . . . . . . . .7.3.1 . . . . . . . . . . . . . . . . . .7.2BAL and ' ! ' . . . . . . . . . . . . .7.3.2 . . . . . . . . . . . . . . . . . .7.3' ! ' . . . . . . . . . . . . . . . . . . . .7.3.3 . . . . . . . . . . . . . . . . . .7.3LID LOCK . . . . . . . . . . . . . .7.3.4 . . . . . . . . . . . . . . . . . . .7.4

Operating Principles . . . . . . . . . . . . . . . . . . . . . . .2.1 . . . . . . . . . . . . . . . . . . . .2.1

Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3.1 . . . . . . . . . . . . . . . . . . . .3.1

PROGRAMMING . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6.1To Enter a Program . . . . . . . . . . . . .6.1 . . . . . . . . . . . . . . . . . . . .6.1To Start a Program . . . . . . . . . . . . . .6.2 . . . . . . . . . . . . . . . . . . . .6.2To Save a Program . . . . . . . . . . . . .6.3 . . . . . . . . . . . . . . . . . . . .6.2

Continued

Iss 1.1

12.4

1CNU12-GB-EFG

Continuation

TOPIC SECTION PAGE

Programming (Continuation)

To Run a Program . . . . . . . . . . . . . .6.4 . . . . . . . . . . . . . . . . . . . .6.2To View a Prgram in Memory . . . . . .6.5 . . . . . . . . . . . . . . . . . . . .6.2To Repeat a Program . . . . . . . . . . .6.6 . . . . . . . . . . . . . . . . . . . .6.3To Stop a Program . . . . . . . . . . . . . .6.8 . . . . . . . . . . . . . . . . . . . .6.3Using Cancel . . . . . . . . . . . . . . . . . .6.7 . . . . . . . . . . . . . . . . . . . .6.3

Recommended Materials . . . . . . . . . . . . . . . . . . . . .8.2 . . . . . . . . . . . . . . . . . . . .8.2Reset, To ~ Cytospin® 7.3 . . . . . . . . . . . . . . . . . . . .7.2

. . . . . .BAL . . . . . . . . . . . . . . . . . . .7.3.1 . . . . . . . . . . . . . . . . . .7.2 . . . . . .BAL and ' ! ' . . . . . . . . . . . .7.3.2 . . . . . . . . . . . . . . . . . .7.3 . . . . . .' ! ' . . . . . . . . . . . . . . . . . . . .7.3.3 . . . . . . . . . . . . . . . . . .7.3 . . . . . .LID LOCK . . . . . . . . . . . . . .7.3.4 . . . . . . . . . . . . . . . . . .7.4

Routine Maintenance . . . . . . . . . . . . . . . . . . . . . . . .8.2 . . . . . . . . . . . . . . . . . . . .8.1Bowl Liner . . . . . . . . . . . . . . . . . . . .8.3.3 . . . . . . . . . . . . . . . . . .8.3Cytoclip™ Slide Clip . . . . . . . . . . . .8.3.8 . . . . . . . . . . . . . . . . . .8.6Front Panel . . . . . . . . . . . . . . . . . . .8.3.1 . . . . . . . . . . . . . . . . . . .8.2Reusable Sample Chamber . . . . . . .8.3.9 . . . . . . . . . . . . . . . . . .8.6Safety Cover . . . . . . . . . . . . . . . . . .8.3.2 . . . . . . . . . . . . . . . . . . .8.2Sealed Head . . . . . . . . . . . . . . . . . .8.3.4 . . . . . . . . . . . . . . . . . .8.2Sealed Head - Base . . . . . . . . . . . . .8.3.5 . . . . . . . . . . . . . . . . . .8.4Sealed Head - Lid . . . . . . . . . . . . . .8.3.7 . . . . . . . . . . . . . . . . . .8.5Slide Clip Support Plate . . . . . . . . . .8.3.6 . . . . . . . . . . . . . . . . . .8.4

Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1.3 . . . . . . . . . . . . . . . . . . . .1.1Save . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.2.8 . . . . . . . . . . . . . . . . . . .5.3Sealed Head - To Close the ~ . . . . . . . . . . . . . . .4.1.2 . . . . . . . . . . . . . . . . . . .4.1

- To Install the ~ . . . . . . . . . . . . . . . .4.1.8 . . . . . . . . . . . . . . . . . . .4.2- To Load the ~ . . . . . . . . . . . . . . . .4.1.6 . . . . . . . . . . . . . . . . . .4.2- To Open the ~ . . . . . . . . . . . . . . . .4.1.1 . . . . . . . . . . . . . . . . . . .4.1- To Remove the ~ . . . . . . . . . . . . . .4.1.9. . . . . . . . . . . . . . . . . . .4.2

Set Acceleration . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.2.11 . . . . . . . . . . . . . . . . . .5.3Set Speed. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.2.1 . . . . . . . . . . . . . . . . . . .5.2Set Time . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5.2.2 . . . . . . . . . . . . . . . . . . .5.2

Continued

Continuation

TOPIC SECTION PAGE

SETTING UP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4.1Disposable Cytofunnels . . . .4.1.5 . . . . . . . . . . . . . . . . . .4.2To Open the Sealed Head . .4.1.1.. . . . . . . . . . . . . . . . . . .4.1To Close the Sealed Head . .4.1.2 . . . . . . . . . . . . . . . . . .4.1To Load the Sealed Head . . .4.1.6 . . . . . . . . . . . . . . . . . .4.2To Load the.Cytoclip . . . . . .4.1.3 . . . . . . . . . . . . . . . . . .4.1To Unload the Cytoclip . . . .4.1.4 . . . . . . . . . . . . . . . . . .4.2To Install the Sealed Head . .4.1.8 . . . . . . . . . . . . . . . . . . .4.2To Remove the Sealed Head4.1.9 . . . . . . . . . . . . . . . . . . .4.2

SPECIFICATION, ACCESSORIES AND CONSUMABLES . . . . . . . . . . . . . . . . . . .11.1Start . . . . . . . . . . . . . . . . . . . . . . .5.4.2 . . . . . . . . . . . . . . . . . . .5.4Stop . . . . . . . . . . . . . . . . . . . . . . .5.4.3 . . . . . . . . . . . . . . . . . . .5.4

TROUBLE SHOOTING . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10.1

View . . . . . . . . . . . . . . . . . . . . . . .6.5 . . . . . . . . . . . . . . . . . . . .6.2

Warning Notices . . . . . . . . . . . . . . . . . . . . . . .1.4 . . . . . . . . . . . . . . . . . . . .1.1Warranty Statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11.1WELCOME . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1.1

Iss 1.1

12.5

1CNU12-GB-EFG

Iss 1.1

12.6

1CNU12-GB-EFG


APPENDIX 1 - METHODOLOGY GUIDELINES

1-1Iss 1.11CNU1X-GB-EFG

Cytospin is a special purpose instrument designed todeposit cells on to glass slides. The instrumentproduces monolayer cell deposition in a defined areaof the slide, using centrifugal force. For most cytologicalspecimens the Cytospin offers significant advantagesin specimen retention, preparation, and standardisation,and ease of specimen evaluation.

Cytological specimens may also be deposited on toslides by techniques such as direct smears or by filtertechniques. While useful with some specimens, bothdirect smears and filter techniques have significantdisadvantages when compared with Cytospinpreparations.

Direct smears consistently produce preparations ofvarying thickness from end to end of the smear. Inaddition, severe mechanical damage may result tomany cells within the preparation. There is also alikelihood of selective cell distribution within thesmear. Cells of different sizes will be deposited in dif-ferent areas of the smear.

Filter preparations, while excellent for cell retention,are technically demanding and time consuming. Inaddition, filter preparations rarely yield slides whichcan be evaluated easily. The cells are seldom in thesame plane as the focus within the microscope, and itis extremely difficult to obtain well stained cells withoutalso staining the filter. For those filter techniqueswhich dissolve the filter, there is a significant risk ofcell loss, in addition to the difficulty and hazards ofusing a volatile and dangerous solvent.

Cytospin preparations effectively circumvent thesedif f icul t ies, and consistent ly produce uni formpreparations of cells which are easily stained andevaluated. In addition, the construction of theCytospin ensures maximum containment of potentiallyhazardous specimens, thereby reducing the risk tolaboratory personnel.

Specimens from body fluids and all body sites can beused for Cytospin preparat ions. The pr imaryrequirements are that the specimen be a cellsuspension, preferably of single cells, and that the

cells are fresh and intact enough to yield diagnosticinformation. With proper application of the generalprinciples of Cytospin operation, consistent preparationsof well preserved cell monolayers should result.

The Cytospin is designed to provide maximumprotection to the operator by completely containingpotentially hazardous specimens. However, Cytospincannot protect the operator during the various stepsrequired to process a specimen prior to using theCytospin. Good laboratory practice requires the useof a biological safety cabinet for all manipulators ofcytological specimens. This includes both the loadingand unloading steps for the Cytospin. Once thespec imen is loaded into the Cytospin sealed head,and the lid is sealed in place, the sealed head maythen be taken outside the biological safety cabinet forspinning in the Cytospin. After the Cytospin hasstopped, the sealed head should be returned to thebiological safety cabinet prior to being opened.

Due to the potential ly infectious nature of thespecimens which may be processed in the Cytospin,the laboratory must establish procedures to ensurethat the instrument is rout inely dis infected.Suggested methods for cleaning and disinfecting ofthe Cytospin and accessories will be found in theCleaning and Maintenance section of the OperatorGuide.

As with al l c l in ical specimens, i t is extremelyimportant to maintain specimen identification. For theCytospin, this means that the slides on to which aspecimen will be deposited must be adequatelylabelled with the appropriate specimen identification.The method of labelling must take into account thesubsequent procedures which will be used. In general,it would be expected that the label may be subjectedto fixation steps and staining procedures. Obviously,a paper label would be inappropriate. Pencil identificationon frosted-end glass slides is the most commonapproach to specimen identification.

APPENDIX 1 - METHODOLOGY GUIDELINESCell Preparation System


The laboratory must also ensure that adequatelabelling is maintained for all containers or devices towhich the specimen is transferred. In use of theCytospin, this may include one or more centrifugationsteps, conducted in a standard laboratory centrifuge.Each new container to which the specimen istransferred must be appropriately labelled. Inaddition, the laboratory must ensure containment ofthe specimen to eliminate potential hazards to thelaboratory personnel. Since most centrifuges do notprovide aerosol containment during operation, anyintermediate centrifugation steps should be conductedin a biological safety cabinet. At the conclusion ofspecimen preparation, all intermediate containers,pipettes, etc., should be disposed of in an appropriatebiohazard container.

Cytological examinat ion always begins with amacroscopic examination of the specimen at the timeit is submitted to the laboratory. This is a crucialexamination, as it provides information which will beused to select processing protocol. The macroscopicexamination is most useful in the hands of anexperienced technologist. Prior experience with aparticular specimen is invaluable in recognisingwhether a given sample is normal or highly suspect,and whether the specimen will be adequate forcytological examination. However, it is usuallyimpossible to determine if a given sample containsabnormal cells from the macroscopic examinationonly. A specimen which should normally be clearshould not be assumed to be abnormal simplybecause it is bloody on arrival in the laboratory. Anynumber of circumstances may produce a differentappearance in a specimen during the collectionprocess.

The macroscopic examination cannot be used as adefinitive test of the specimen. It does serve tosupport the eventual diagnostic assessment, butmore importantly, it provides the information whichwill allow the technologist to choose a specimenpreparation protocol. A complete macroscopicexamination may include:

1 Record of specimen origin - precise anatomical site.

2 Quantity of specimen.

3 Specif ic Gravity ( i f specimen is fresh andunfixed).

4 Odor, if present.

5 Gross characteristics.

Gross charac te r i s t i cs descr ibe the phys ica lappearance of the specimen. Important parametersare the color of the specimen, its viscosity, andwhether the specimen is homogeneous or containssolid tissue fragments.

The gross examination will also determine if thespecimen is fresh or if it has been fixed prior todelivery to the laboratory. In general, it is preferableif all cytological samples are submitted to the laboratoryin the fresh state. However, in many cases, due totransport distances or time constraints, the specimenwill be fixed prior to submission. This must be notedduring the gross examination, as fixation may affectseveral of the parameters recorded during the grossexamination. Prior fixation may also constrain thesubsequent processing of the specimen. Fixationand its effects will be discussed in a subsequentsection of this paper.

Successful operation of the Cytospin requires knowledgeof the number of cells present in the sample. Whilethe experienced technologist will achieve reasonableresults by estimating the cell number, less than optimalpreparations sometimes result from such estimates.It is highly recommended that all specimens areexamined specifically to determine cell numbers.Visual appearance alone is often confusing, sincespecimen turbidity may be the result of cell debris,suspended lipids, or other non-cellular materials. Insuch cases, a direct determination of cell number isnecessary to ensure proper Cytospin preparations.

Samples which contain ‘average’ cells, that is cellswith an approximate diameter of ten to twelvemicrons, produce excellent Cytospin preparations at

cell densities of one million cells (1x106) per ml.



Iss 1.11CNU1X-GB-EFG1-3

Specimens containing large cells require lower cellconcentrations, and specimens with tiny cells, cello rgane l l es , o r bac te r i a , may requ i re h ighe rc o n c e n t r a t ions. The absolute concentrat ionrequired wi l l be somewhat dependant on theprocessing methodology employed. As a generalrule, the concentration chosen should be such thatthe cells within the sample have adequate space tospread into a monolayer on the slide surface, withminimal overlap, or piling up of cells. Ideally, theconcentration should be high enough that there is nottoo much space between cells. Having sufficientconcentration of cells speeds up evaluation of thepreparation, since little time will be lost in searchingfor cells to evaluate.

A quick method for approximating the number of cellspresent in a sample is to place a single drop of thesample on a slide and cover with a 24 x 50 mmcoversl ip. By lower ing the condenser of themicroscope, or by closing the microscope condenserdiaphragm, the unstained cells can be seen ( althoughdetail will not be seen). Using the 10 X objective,scan the field and pick an area that appears aboutaverage for the entire slide. The cells will mostly likelynot be evenly spread, which is why it is necessary toselect an average area. Now switch to the 40 Xobjective. You may also need to open the diaphragmor raise the condenser slightly.

Count the cells seen in the field. It is not necessary tocount every cell - an approximation will do. Refer tothe illustration Cells/X 40 Field as a Guideline to theNumber of Drops of Cell Suspension/CytospinSample Chamber. This count can be used toestimate the number of drops of the cell suspensionrequired for a Cytospin preparation. To determine thenumber of cells being used, multiply the number ofcells counted by 38. Divide the number of cellscounted into 60. The quotient equals the number ofdrops that should be added to the Cytospin samplechamber, though the total volume should not exceedthe 0.5 ml capacity of the chamber. This gives thetotal number of cells applied to the Cytospin funnel foreach drop of suspension used. While this techniquefor estimation of cell number is an approximationonly, it does provide an excellent control of Cytospinpreparations.

A second method of cell number determination is bymanual counting of cells in a hemocytometer. This is

a device which defines a precise volume between aspecial glass slide and a coverslip. By counting thetotal number of cells within this volume, the cell con-centration within the specimen can be accuratelydetermined. While more accurate than simply using acoverslip on a standard slide , the extra precision ofthis method is not usually required for successfulCytospin preparations.

A third method for determination of cell number is byuse of a cell counter of the electronic volume sensingtype (Coulter counter). This instrument can provide aprecise evaluation of a cell sample. It does requiresufficient amount of sample however, which may notalways be available. It is common to determine thecell number of specimens using this instrumentationin the hematology laboratory. Any specimenobtained from the hematology laboratory mayinclude cell number (or concentration) information.

It is important to recognise that samples which arequite concentrated should be handled carefully. Forexample, if the specimen is so concentrated thatonly a single drop may be required, the addition of asecond drop will double the cell concentration. It ispreferable to work with specimens that are diluteenough that five or six drops are required for thepreparation. With such a specimen, the addition of onemore drop will not be as likely to result in overlappingcells in the final Cytospin preparation.

While it is common to discuss Sample sizes in termsof ‘drops’, it is important to realise that drop size willbe dependant on the type of pipette used to transferthe specimen. As an example, a Falcon 3 ml transferpipette will dispense approximately 0.5 ml in 15drops. A six inch glass Pasteur pipette will dispense0.5 ml in 20 drops. It is advisable to standardise on asingle pipette type for all cytological preparations,otherwise ‘drops’ wi l l be a meaninglessmeasurement.

Many cytological specimens arrive in the laboratoryas relatively large fluid volumes, many with relativelyfew cells. Such specimens must obviously beconcentrated prior to use of the Cytospin. Suchconcentration requires the use of a general laboratorycentrifuge. The amount of the specimen submitted



The large circles represent cells in one drop of cell suspension, either of unprocessed cell specimen orpreferably of centrifuged cell concentrate, that have been spread under a 24 x 50 mm cover glass andviewed under a 40X objective. Although drawn smaller than they appear microscopically, the cell and fielddiameters are proportional to one another at a 50:1 ratio. Simply match the microscopic field with its closestcounterpart here and use the number of drops so indicated.

1 Drop 2 Drops 3 Drops

6 Drops5 Drops4 Drops

12 Drops 20 Drops

CELLS/40X FIELD AS A GUIDELINE TO THE NUMBER OF DROPS OF CELLSUSPENSION/CYTOSPIN SAMPLE CHAMBER




will determine the size centrifuge tubes which will benecessary. In some cases, the original specimenmay need to be split between many tubes. As anexample, if a centrifuge is available which can hold 50ml tubes, and the total amount of specimen is 100 mlor less, then two tubes can concentrate the entirespecimen. Should the specimen amount to 150 ml,then four tubes would be required. Remember,centr i fugation wil l also wi l l also require equalnumbers of tubes. By adding more tubes it is possibleto concentrate specimens which are quite large involume. To sediment the cells, the centrifuge shouldbe spun at 2000 to 3000 r.p.m. for 10 to 20 minutes.Avoid spinning at excessive speeds. This will onlydamage cells, and pack them into such tight buttonsthey will be difficult to process further. Centrifugeswith swinging buckets will generally require slightlyhigher speeds than angle headed centrifuges.

After conventional centrifugation, any cells present inthe sample would appear as a packed button in thebottom of the tube. The clear supernatant above thecell button should be carefully aspirated and pouredoff, leaving a volume of fluid approximately equal tothe volume of the packed cell button.

The fluid that is aspirated or poured off may bediscarded, using any common technique to render itharmless (sterilisation, fixation etc.). This fluid shouldonly be discarded after it is determined that the cellswithin have indeed been retained.

The packed cell button in the bottom of the centrifugetube should be thoroughly mixed with the residualfluid which was not removed. This is done either byuse of a vortex mixer, or by gentle agitation of thetube. The result is a concentrated cell suspension,suitable for cell number determination, and subsequentpreparation with the Cytospin.

Cytology specimens often are submitted to thelaboratory with a cell concentration that is too high forCytospin preparations until diluted. Such specimensare common from bone marrow, lymphoid aspirates,and many fine needle aspirates. These concentratedspecimens should first be evaluated for cell number,using any of the previously described techniques.The specimen should then be diluted to an approximatecell concentration by addition of a balanced electrolyte

solution. It is important to use a fluid that has a properosmolarity, in order not to introduce structuralchanges in the cell sample. Simple solutions ofSodium Chloride (0.9% saline) are unsuitable as diluents- they produce rapid changes in nuclear chromatinand interfere with subsequent cytological evaluation.

Suitable di luents are those commonly used inhematology laboratories, such as Polysal, PolyonicR-148, Plasma-Lyte, Isoton, Tisu-u-Sol, or Normosol.Many of the solutions commonly used in tissueculture laboratories are also suitable diluents, such asGey’s balanced salt solution, Earle’s balanced saltsolution, or Hank’s balanced salt solution. In manycases, i f the cell suspension submitted to thelaboratory has been col lected in one of thesediluents, or has undergone significant processing insuch salt solutions, it may be advisable to add someprotein to the diluent. Either human serum or asolution of bovine serum albumin may be used. Theusual concentration of these solutions is 1 to 30percen t . The p ro te in so lu t ions o f ve ry h ighconcentration are usually used by dropwise additionto the sample just prior to or during loading of theCytospin funnels.

The sample chambers hold a maximum of 0.5 ml ofspecimen and should hold no less than 0.1 ml. It ismandatory that no more than 0.5 ml of sample isplaced in each sample chamber. Additional samplewould simply be thrown to the top of the chamberduring Cytospin operat ion, and could not bedeposited on the slide. It is recommended to load thesample chambers after they have been assembledand inserted into the sealed head. The design of thechamber assemblies and the sealed head ensuresthat the sample chambers tilt in such a manner thatthe specimen will not contact the slide or the filtercard prior to starting the Cytospin. The specimenmust never contact the slide or filter before theCytospin is started. The operator must be carefulduring loading not to forcibly inject the sample into thesample chamber. The sample should be eased intothe sample chamber slowly, allowing ample opportunityfor air to be displaced by the sample. Cytofunnel™disposable sample chambers are available for addedsafety and convenience.



For concentrated cell suspensions which require onlyone or two drops of sample to obtain the correct cellconcentration, it may be sometimes necessary to addadditional diluent to bring the total volume in thesample chamber up to 0.5 ml. This addition can bedone in the chamber as the samples are loaded.However, this requires care to avoid forcing sampleinto the slide/filter area and it is recommended that‘thick’ specimens are diluted prior to being loaded intothe Cytospin.

During loading of the sample chamber, the sampleshould be deposited directly in the bottom of thesample chamber. Avoid dripping the sample downthe side of the sample chamber. Should sample bedeposited on the walls of the chamber, rinse downwith a drop of diluent. The object is to ensure that all ofthe sample is in the bottom of the sample chamber.

The speed of operation of the Cytospin is dependanton the size of the cells or particles to be deposited onthe slides. In general, average cellular specimenswill require a speed of approximately 1000 r.p.m.Very large cells and fragile cells may require a slowerspeed, such as 500 to 800 r.p.m. Specimens consistingof tiny objects such as bacteria may require muchhigher speeds, approaching 2000 r.p.m.

Time of Cytospin operation is also related to specimentype and to subsequent preparative steps. For mostcytological preparations, it is desirable to avoid anypossibility of air drying of the specimen. Thereforethe time used for Cytospin operation is kept as shortas possible, such as 3 to 4 minutes. For hematologyand microbiology specimens which often are air driedprior to further processing, a longer time is used,often approaching ten minutes.

An appropriate Cytospin time will ensure fluid absorption.The cells on the slide should have a thin layer of fluidon their surfaces. Occasional specimens may be toothick to completely absorb in the filter paper during anormal time and speed setting. Such specimens mayrequire special processing. An example is jointaspirations which contain hyaluronic acid, givingthem a thick consistency. This can be reduced byadding a small amount of the enzyme hyaluronidaseto the sample prior to operation of the cytocentrifuge.

After the Cytospin stops, the specimens should beremoved as quickly as possible. The lid of theCytospin is opened, and the sealed rotor is removedand taken to the biological safety cabinet. The lid isopened, and the ind iv idua l sample chamberassemblies are removed from the rotor. For cytologyspecimens, the assemblies are laid on flat towelling,and the clips released. The chamber assembly is liftedstraight up on occasions where liquid remains, allowingany residual fluid to flood the slide. If there is aconsiderable amount of residual fluid, wait until someof it evaporates. However, do not allow the specimento dry. Just before drying begins, place the slide infixative, or spray with Cell-Fixx™.

For specimens which do not contain excess fluid,quickly remove the sample chamber, remove theslide, and immediately place into fixative. (This is bestachieved by easing the slide into the fixative, so asnot to disturb the deposited cells).

Specimens intended for air drying techniques, suchas hematology specimens and some microbiologyspecimens, are handled somewhat differently. Inthese specimens, it is undesirable to have residualfluids flood the slide. The chamber assemblies arestood upright on the towelling prior to unclamping.Then the clips are released and the chamber pulledaway from the slide. Residual fluid is retained in thesample chamber, and does not f lood over thedeposited cells. The slides resulting from thisprocedure are then completely air dried prior tofurther processing.

After staining, the specimens can be evaluated toassess the preparative technique.

The ideal result is a monolayer of cells with minimaloverlap, yet sufficiently concentrated that one doesnot have to search for cells in the preparation. Thecells should display excellent morphology. Thereshould be no evidence of stretching, or tearing of theperiphery of cells. Such artefacts indicate excessivespeeds or times of cytocentrifugation. In excellent




preparations, there will be flattened nuclei withdistinct chromatin patterns. Some cell types, such ascolumnar epithelial cells should retain their typicalcolumnar morphology. Distortion of their columnarshape ind ica tes excess ive speed o r t ime o fcytocentrifugation.

Occasionally one will see a specimen that has apattern of cell deposition around the periphery of thedeposition spot, with a loss of cell in the center. Thiseffect is due to an excess amount of residual fluid inthe center of the cell deposition area when thespecimen is fixed. Because the cells in the center ofthe area are quite wet, they wash off the slide as it isimmersed into the fixative. The solution to thisproblem is to allow longer time for the slide to dryprior to fixation, and to be exceptionally gentleduring immersion of the slide into the fixative.

Fixation is used to preserve cell samples, to renderthem more easily stained, and to produce characteristicpatterns of cell structure which are used to distinguishcell types. Cells continue their natural living processesafter being removed from body sites. Since they nolonger have their normal blood supply and othersupporting environment, they will begin to degenerateas they run out of required nutrients and gases andbegin to build up waste products. As these eventscontinue, the cell activates internal repair mechanismsthat eventually result in the cell digesting itself. Thisis called autolysis. The rate at which autolysisprogresses is different for different cell types, butdoes mean that samples should be processed asquickly as possible. Autolysis can be slowedsignif icantly by refrigeration, and samples may beheld for some period of time at refrigerator temperature.Where practical however, specimens should be fixedor processed as soon as possible.

Fixatives are chemical agents that both kill the celland stabilize its structure. The ‘killing’ also inactivatesmany of the enzyme systems of the cell, particularlythose associated with autolysis. Fixatives thereforealso function as preservatives, and well fixed cytologicalsamples essentially last indefinitely. Many specificchemicals can be used as fixatives, and each hasspecific properties that are desirable for certain typesof study. These fixatives include those that produce

chemical cross-links within the tissues, such asformalin, and those that precipitate cellular componentssuch as the alcohols. By far the most common fixativeused in cytological studies is alcohol. Alcohol producesdistinct nuclear chromatin patterns, and also serves toremove water from the cells. Cell-Fixx spray fixa-tive is an example of an alcohol based cytological fix-ative which also contains Carbowax (Carbowax isa registered trade name of Union Carbide).

A disadvantage of alcohol fixatives is that theyevaporate quickly, and therefore there is always therisk of permitting specimens to dry out. To avoid this,many laboratories use Saccomanno fixative, whichis a mixture of 70% ethyl alcohol and 2% Carbowax(polyethylene glycol). The Carbowax in this mixtureforms a coating over the specimen, helping protect fromthe effects of drying. The Carbowax is soluble in water,and so is dissolved in subsequent staining steps.Commercial versions of this fixative are available(Thermo Shandon Cytospin Collection Fluid).

When specimens must be transported over considerabledistances, or when they cannot be processedimmediately, it is advisable to fix with alcohol or withSaccomanno type fixative. This is done by adding anequal volume of fixative to the sample. If the specimenis large in volume, the sample should be centrifugedto concentrate the cells, and then fixed. Immediatelyafter adding the fixative to the sample, the sampleshould be vigorously agitated to suspend the samplewithin the fixative.

Occasionally samples will be received that have beenfixed in some other fixative such as formalin. Thesewill have a different nuclear and cytoplasmic appearance ifprocessed without exposure to alcohol. Such specimenscan be concentrated, the formalin poured off, andthen re suspended in Saccomanno-type fixative. Theresult will be a specimen that is reasonably similar tothose fixed in the alcohol fixative alone.

Fixation makes cells more rigid, and less able tospread when placed in the Cytospin. Specimens thathave been fixed prior to depositing on slides willrequire slightly higher speeds and longer times toachieve the same degree of spreading as seen inunfixed specimens. Occasionally the laboratory willbe asked to prepare specimens that have been heldin fixative solutions for extended times. These specimensmay be so rigid that it is difficult to get them to flattenon the slide. The addition of a small amount of glycerol to



the specimen, allowing some ‘soak’ time, followed byuse of the Cytospin, will usually result in a reasonablepreparation.

Many laboratories prefer to fix all specimens duringpreparation. The usual protocol is to concentrate thespecimen, then re suspend in an equal volume ofSaccomanno-type fixative. For samples that must bediluted, the diluent can be fixative. These fixationsteps are generally done just before adding the sampleto the Cytospin sample chamber assemblies.

Whether the specimen has been f ixed beforedeposition on slides or not, immediately after removingthe sl ides from the Cytospin, they should beimmersed in 95% alcohol to complete fixation anddehydration. Since the cells will still be wet, and willnot have become totally bound to the glass slide, usecare in this transfer. (Ease the slide into the fixative inthe container). Many complaints of poor cell capturecan be traced to lack of care in this step of theprocedure.

Successful application of the Cytospin requires thatcells adhere to the glass slides. For many routineapplications, it is sufficient to use clean slides. Slidesmay be cleaned using alcohol. The increasing use oflong staining techniques, such as immunostaining,may require additional steps to ensure adhesion ofsamples. Slides may be coated with Poly-L-Lysine(Sigma) or with aminosilane (Sigma). The use of anyof these adhesion methods will increase cell retentionand reduce the incidence of ‘floater’ in the subsequentstaining baths.

The majority of the procedures previously discussedapply to cytological specimens. However, there are anumber of specific specimen types which requirespecial processing. Often, the cytology specimen willcontain clots, fibrin webs, or tissue fragments. These

will all interfere with he Cytospin preparations. Smallfloating clots and fragments, should be removed withforceps. These may be saved for ce l l b lockprocedures. Fibrin webs are generally too friable tobe removed intact. These are most easily removedby twirling a glass or wooden rod in the specimen.The fibrin web is wound onto the rod, and in theprocess, many of the trapped cells will float free.After winding on the fibrin, it is pressed gently againstthe side of the container to squeeze out as much ofthe trapped fluid and cells as possible. As with all cellmanipulation techniques, it is important to be gentleto avoid excessive cell damage.

Cytological samples may contain considerablequantities of mucus. This a very thick mass that isdifficult to dilute or concentrate, and becomes arubbery mass on fixation. Such specimens should beprocessed prior to fixation. A common way to breakup mucus is to mix wi th an equal part ofSaccomanno-type fixative and immediately blend in asmall blender. Three to five seconds is usuallysufficient. The blending procedure should be done ina biological safety cabinet. After blending, thesample should be homogenous and non-stringy, andcan be deposited using the Cytospin. Certainchemicals also react with mucus to produce liquefaction,such as acetylcysteine (Boccato, 1981). A commercialproduct of this type is Mucolexx, which contains notonly a mucolytic agent but Saccomanno-type fixativeas well.

Bloody or serosanguineous specimens may containso many erythrocytes that examination of cytologicalpreparations is difficult, and when the specimen isdiluted sufficiently to obtain monolayer preparations,the cells of interest are difficult to locate. Red bloodcells may be removed by gradient centrifugation or byvarious lysing procedures. Lysing techniques arecommonly used for leukocyte counting on cel lcounters which employ sensing orifices. A number ofcommercial lysing reagents are available and providecomplete destruct ion of erythrocytes. Lysingreagents may also damage some cells of cytologicalinterest, and care must be used in their use. Thelarge amounts of hemoglobin released from the redcells may interfere with subsequent staining. It canbe removed by several washing steps using aconventional centrifuge.

Gradient centr i fugat ion is based on a densitydifference between red cells and other cells within a




specimen. Commercial gradients are available, andare based on mixtures of Ficoll and Hypaque. Thesample is layered onto the gradient in a centrifugetube and then the tube is centrifuged. The red cellswill migrate through the gradient, and will also behemolyzed. The remaining cells will stay on top ofthe gradient, and can be removed for subsequentprocessing.

The primary difference between cytological andhematological preparations is the routine use of driedair preparations in hematology. In most cases, hema-tology will also have available a specific cell countderived from an electronic volume sensing instru-ment. This permits a defined solution and allows pre-cise control of cell number on the final Cytospinpreparation.

Hematolgical samples are routinely diluted to obtainsamples of the correct cell concentration. The diluentused is commonly one of the balanced salt solutions.it is advisable to add a small quantity of serum albumin(preferably bovine for safety) to the sample toincrease adhesion of the cells to the slide, and toavoid the deleterious effects of the high concentrationas the diluent evaporates during the drying of theslide.

Since hematology specimens are to be air dried, theyshould not be flooded with any residual liquid afteruse of the Cytospin. For hematology, after thechambers are removed from the Cytospin sealedhead they are stood upright. This drains any residualfluid back into the sample container. The clips arethen removed, and the slide separated from thesample container and filter. The slide is then airdried. As with all air dried preparations, the morerapidly drying occurs, the better. Drying may beaccelerated by gentle heating of the slides.

Specimens derived from urine are typically highvolume fluids with few cells or particles. Thesespecimens must be concentrated by conventionalcentrifugation prior to cytological examination. Oftenurine samples contain particles which are not cellular

but are precipitated phosphate salts. It is oftensuggested that one can dissolve these particles byacidifying the specimen with a few drops of aceticacid. While this will cause the salts to dissolve, aceticacid is also a classic fixative. It will cause chromatincondensation in the cell nuclei, and will also producesignificant cell swelling. Acetic acid is a componentof Carnoy’s fixative. If acetic acid is used, its effectsmust be accounted for in subsequent evaluation ofthe specimen.

Many microbiology samples are quite similar tocytological samples. They will contain cells and themicrobiologist is interested in the association ofbacteria or viruses with the cells. The Cytospin canalso be used to directly deposit samples of bacteriaonto slides. The advantages of this use is that thedeposited bacteria are generally more concentratedthan after simple smearing, and they are all located ina defined area of the slide. Due to the small size ofthe bacteria, the Cytospin is generally operated atspeeds of 2000 r.p.m. for five to ten minutes.

Many of the techniques used for localization ofviruses or bacteria require the use of fixatives otherthan alcohol based ones. In general, techniquesusing immunostaining or nucleic acid probes willspeci fy the use of aldehyde f ixat ion, such asformaldehyde (paraformaldehyde) or gluteraldehyde.These do not affect the operation of the Cytospin,since they are applied to the slide after the cells,bacteria, or viruses are deposited on the slide. It isimportant to use a slide adhesive, preferably Poly-L-Lysine or aminosilane for these preparations.

.

This section is designed to provide general guidelines for the use of the Cytospin in hematology and clinicalmicroscopy.

The Cytospin has a wide range of applications in clin ical microscopy, some of which are shown below.



USES CLINICAL APPLICATIONS1 Romanowsky-stained cytology of CSF and Evaluation of possible infection or presence of

other body fluids. malignancy.2 Gram stain and other special stains of CSF Detection of infectious agents:

and body fluids. characterization of malignant cells.3 Slide preparation from ficoll-hypaque cell Provides slides for morphology, cytochemical staining

isolates. (e.g. myeloperoxidase, non-specific esterase), and immunocytochemical or immunofluourescent assays (e.g.TdT). Used to characterize leukemias and lymphomas.

4 Cell surface and cytoplasmic marker studies Classification of leukemias and lymphomas.using monoclonal antibodies.

5 Urine eosinophis. Evaluation of drug-induced nephritis, allergic cystitis, and renal transplant rejection.

6 Urine hemosiderin. Confirmation of severe intravascular hemolysis and chronic iron overload.

When first setting up the Cytospin for use, it is helpfulto establish a standard procedure, i.e. the amount ofspecimen/slide, r.p.m. and minutes centrifuged, andmaximum number of white blood cells/ìl and red bloodcells/ìl, above which dilutions would be necessary.

1 Determine the red cell and white cell count of thesample according to established methods.

2 Using a ‘standard’ amount of specimen, (e.g. 5 dropsor approximately 0.25 ml) make serial dilutions ofa highly cellular specimen to determine the maximumnumber of white blood cells and red blood cellsthat can be present in a specimen before dilutionsare necessary. Some tests may require more cellularslides than others.

3 Experiment with a range of r.p.m. to see whichgives the most desirable morphology for the procedureinvolved. Establish the minimum number of minutesrequired to spin the entire standard amount of

specimen onto the slide. Experiment with the ‘high’and ‘low’ acceleration settings to see if there isany difference in morphology on the cells.

4 The following is an example of a standard procedurefor preparing slides for a white cell differentialwith Wright stain:

Use 5 drops of specimen/Cytospin chamber.Dilute sample to obtain a white cell count less than500/ìl and a red cell count less than 5,000/ ì l .Centrifuge at 700 r.p.m. for 5 minutes.

Dilution Chart for WBC DilutionWBC COUNT DILUTION

0 - 500 None501 - 1000 1/2

1001 - 1500 1/31501 - 2000 1/42001 - 2500 1/52501 - 3000 1/6

If the red cell count which results from the aboveWBC dilution is greater than 5,000/ìl, calculate thefurther dilution necessary to bring the red cel lcount below 5,000/ìl. Use this dilution regardless ofthe fact that the white cell count may be very low.

5 Assemble the sample chamber as per theinstructions in the Operator Guide. Be sure toalign the outlet hole of the sample chamber exactlyinside of the hole in the filter card. The edge of thesample chamber must not touch the sample card orthe cell yield will be greatly reduced.

6 Always place the clip assembly into the Cytospinhead before adding anyth ing to the samplechamber. Check that the assembly pivots freely inthe metal support plate.

7 Add one drop of 30% albumin to the bottom of thesample chamber, then add the standard amount ofdiluted fluid. Cap the sample chambers. Thealbumin helps to preserve the cells and keep themin contact as they are deposited onto the slide.(Note: albumin may quench fluorescence andshould not be used when immunofluourescentassays are to be performed).

8 Lock the lid down on the base, transfer the sealedhead to the Cytospin, and close and lock the lid.Never snap the lid of the Cytospin head down ontothe bowl while the assembly is sitting on the driveshaft, as this may damage the shaft.

9 Program the Cytospin to the standard number ofr.p.m. and minutes and start the unit. Fragile cellsmay require a lower r .p.m. set t ing and lowacceleration to maintain morphology.

10When the alarm has ended, remove the sealedhead and transfer it to a hood before opening.

11Check the sample chambers to see if all of thespecimen has spun onto the slide. Make sure thatany remaining specimen in the chamber does notflow onto the slide as it is removed from the clip.Should this occur, the cells will not remain spreadout on the slide and may therefore overstain.




1 Use one drop of 30% albumin at the bottom of theCytospin chamber.

2 Do not make a push smear, even when the cellcounts are high. Large malignant cel ls aredifficult to identify on push smears because theymay aggregate at the feather edge or stain darklyin the thick portion of the smear.

3 Centre the outlet port of the sample chamberexactly inside of the hole in the filter card.

4 Do not let unspun specimen wet the slide when thechamber is disassembled.

5 If fibrin strands or other contaminated materials arepresent, they may clog the filter card and preventabsorption of the specimen. Better slides maysometimes be obtained if an aliquot of the spec-imen is first diluted in saline, centrifuged, andthen the cells are re suspended in saline to the orig-inal volume.

6 If synovial fluid is extremely viscous, a smallamount of hyaluronidase may be added toliquefy the sample before processing.

7 I f a f luid is clotted, Cytospin sl ides may beprepared from a suspension of the clotted materialas well as from the un clotted fluid to increase thepossibility of detecting malignant cells.

8 Avoid bacterial contamination of the albumin orsaline diluent. Report the presence off bacteria onthe slides only when they are also present inneutrophils. Make sure that slides prepared withreagent only are free of bacteria.

These guidelines outlining the Use of the Cytospin forHematalology and other Clinical Microscopy

Specimens have been graciously prepared by:Nicky Sherwood, MT (ASCP)

andJoanne Cornbleet (MD)

Clinical Hematology LaboratoryStanford University Medical Center.

Iss 1.12-1

1CNU2X-GB-EFG

PRODUCT RETURN SAFETY DECLARATIONPart 1 DECONTAMINATION CERTIFICATE

Any instrument or part of any instrument must be clean before being returned, and where necessary accompanied by acompleted Decontamination Certificate. Should the instrument or any part of it be received in an unclean condition, or ThermoShandon Limited consider it to be a hazard, the instrument or part will be returned unrepaired at the expense of the customer.

It is important that the certificate is forwarded by post or fax, and a copy attached to the exterior of the container. Containers will notbe opened until the company is in possession of the required certificate.

This form MUST be completed by the customer and NOT a Thermo Shandon or distributor employee.

If an instrument or part is to be returned to THERMO SHANDON LIMITED, please note the following:-

1 If the instrument or any part of it has been exposed to, or been in contact with potential pathogenic or radioactive material, it isessential that it is decontaminated.

2 Set procedures are laid down in the European Health and Safety Directives for decontamination. To avoid any misunderstanding,we request that all instruments or parts returned to us must be accompanied by a certificate stating the following:

We certify that this (Model) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Serial No . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

· has not been exposed to pathogenic, radioactive or other hazardous material and has been cleaned, OR

· has been decontaminated and cleaned (if exposed to the above) according to approved procedures, following exposure to:

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Signed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Position . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Name (Block Capitals) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Company or Organisation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Full address . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Part 2 Guidelines for Returning Instruments

Please use the checklist below to ensure that the instrument being returned is ready for collection.

· All reagents / wax removed from instrument, including vapour traps (if applicable) . . . . . . . . . . . . . . . . . . . . . . . . . . .

· Accessories are secured / itemised . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

· Instrument has had transit clamps fitted as per operator guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

· Instrument is packed in original packaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .YES / NO

RMA NUMBER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

CARRIER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

FOR ATTENTION OF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Thermo Shandon Limited, 93-96 Chadwick Road, Astmoor, Runcorn, Cheshire, WA7 1PR, United KingdomTel: +44 (0) 1928 566611; Fax: +44 (0) 1928 565845; www.thermoshandon.com

· Has the instrument been used for work with human, or animal, Transmissible Spongiform Encephalopathies, e.g.Creutzfeld-Jacob disease, Scrapie or BSE?

YES / NO

If yes, please contact Thermo Shandon Service before taking any further action.

2-21CNU2X-GB-EFG Iss 1.1

issue 1 operator guide - marshall scientific...3.1.6 cytospin ® 3 is a class 1 instrument as...

Documents