ser serialdilutions

7/24/2019 Ser SerialDilutions

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Serial DilutionsSerial Dilutions

MLAB 1335MLAB 1335

Immunology/SerologyImmunology/SerologyTerry KotrlaTerry Kotrla


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IntroductionIntroduction

Many of the laboratory proceduresn!ol!e the use of dlutons"

It s mportant to understand theconcept of dlutons# snce they are ahandy tool used throughout all areas ofthe clncal laboratory"

These dlutons ha!e to be consderedas they ma$e a %uanttat!e d&erencen 'hat s gong on"


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A serial dilution s any dluton'here the concentraton

decreases by the same quantityin each successive step"

Seral dlutons are mutplcat!e"


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What Does ThisWhat Does This

Mean??Mean?? If a soluton has a 1/1( dluton the

number represents 1 part of the

patent sample added to ) parts ofdluent" So the !olumes used 'ould be 1(*

1+ )"Ths represents 1 part patent

sample added to ) parts of dluent"


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DilutionsDilutions

If a 1/, dluton of the stoc$soluton s made follo'ed by a 1/-

dluton 'hat s the .nal dluton"The .nal dluton s 1/, 0 1/- +

1/,

These type of dlutons are trc$erThese type of dlutons are trc$erand not used !ery fre%uently nand not used !ery fre%uently n

the clncal lab"the clncal lab"


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Doubling DilutionsDoubling Dilutions

2oublng dlutons4 are !erypopular"

Ths s a seres of dlutons"6ach success!e tube 'll theamount of the orgnal

concentrated soluton" If ths s done - tmes ths s 'hat

you 'ould end up 'th


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Doubling Dilution 6Doubling Dilution 6

TimesTimes 1st dluton + 1 /7

7nd dluton + 1 /7 0 1 /7 + 1/

3rd dluton + 1/ 0 1 /7 + 1/, th dluton + 1/, 0 1 /7 + 1/1-

5th dluton + 1/1- 0 1 /7 * 1/37

-th dluton + 1/37 0 1 /7 + 1/- Ths results n a seres of dlutons# each

a doublng dluton of the pre!ous one


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Dilution FactorDilution Factor

The dluton factor s the .nal uses theformula !olume/al%uot !olume"

68AM9L6 :hat s the dluton factor fyou add ("1 mL al%uot of a specmento )") mL of dluent;


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PracticePractice

9roblem :hat s the dlutonfactor 'hen ("7 mL s added to

3", mL dluent;


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Set Up The ProblemSet Up The Problem

dluton factor + .nal!olume/al%uot !olume

("7 >3", + "( total !olume

"(/("7 + 17( dluton


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Problem ContinuedProblem Continued

?emember that seral dlutons areal'ays made by ta$ng a set

%uantty of the ntal dluton andaddng t success!ely to tubes'th the same !olume"

So each success!e dluton 'ouldbe multpled by the dlutonfactor"


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Problem ContinuedProblem Continued

So n the abo!e problem allsuccess!e tubes 'ould ha!e 3", mLs

of dluent"@ou 'ould then transfer ("7 of the

ntal dluted sample nto the ne0ttube# m0 transfer ("7# m0 and so on"

If you had tubes 'hat 'ould be the.nal dluton of tube ;


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Solving the Problem Solving the Problem

!Calculate DF o" tube #!Calculate DF o" tube #TubeTube 11 77 33

Al%uotAl%uot ("7("7 ("7("7 ("7("7 ("7("7

luenluen

tt3", 3", 3", 3",

MathMath/("/("

771/7(01/7(1/7(01/7( 1/((01/71/((01/7

((1/,(((01/7(1/,(((01/7(

lutolutonn

17(17( 1((1(( 1,(((1,((( 11-(#(((11-(#(((


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Solving the ProblemSolving the Problem

r f you smply 'anted to $no'r f you smply 'anted to $no'

the dluton of the .nal tube youthe dluton of the .nal tube you

could Cust multply them togethercould Cust multply them together 1/7( 0 1/7( 0 1/7( 0 1/7( + 11-(#(((


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$e%t$e%t

:hat f 'e added the ndcator system:hat f 'e added the ndcator system

to the test system;to the test system;

@ou must create addtonal ro's# f you@ou must create addtonal ro's# f youare sho'ng your 'or$# to ta$e ntoare sho'ng your 'or$# to ta$e nto

consderaton the dlutonal e&ect thatconsderaton the dlutonal e&ect that

'll ha!e on the dluton"'ll ha!e on the dluton"

:hen an ndcator s added t s:hen an ndcator s added t sD?ITIDAL to remo!e an al%uot fromD?ITIDAL to remo!e an al%uot from

the last tube"the last tube"


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&dding Indicator Cells&dding Indicator Cells

TubeTube 11 77 33

lutoluto

nn

17(17( 1((1(( 1,(((1,((( 11-(#(((11-(#(((

?BDs?BDs ("7("7 ("7("7 ("7("7 ("7("7

EolumEolumee

3", 3", 3", 3",

FF /1/1

MathMath 07(07( 0((0(( 0,(((0,((( 01-(#(((01-(#(((

luto 1,(1,( 1,((1,(( 137#(((137#((( 1-(#(((1-(#(((


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:hen performng seral dlutons n theclncal laboratory one must $no' the

dluton of each tube so a chart sal'ays created to ndcate the dlutonof each tube"

If you $no' the dluton factor you

multply each success!e tube by t andcan easly determne the concentratonof each tube to create your chart"


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TitersTiters

TIT6?S are reported out as therecprocal of the last tube g!ng a

post!e reacton" So f tube 7 'as the endpont#

the dluton s 1,(( the tter s

reported out as ,((/1+ ,(("


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'ther &pplications'ther &pplications

Sometmes 'hen analyGng aSometmes 'hen analyGng a

blood sample the results areblood sample the results are

abo!e the lnearty Hor accuracyabo!e the lnearty Hor accuracyof the machne"of the machne"

:hen ths happens the blood:hen ths happens the blood

must be dluted# retested# thenmust be dluted# retested# thenthe result multpled by thethe result multpled by the

dluton factor"dluton factor"


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(%ample(%ample

A blood glucose of ,(( mg/dL 'asA blood glucose of ,(( mg/dL 'as

obtaned"obtaned"

Accordng to the manufacturerAccordng to the manufacturerthe hghest glucose result 'hchthe hghest glucose result 'hch

can be obtaned on ths partcularcan be obtaned on ths partcular

nstrument s 5(( mg/dL"nstrument s 5(( mg/dL"The sample must be dluted"The sample must be dluted"


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(%ample(%ample

The serum 'as dluted 11( andThe serum 'as dluted 11( andretested"retested"

The result s ,( mg/dL"The result s ,( mg/dL"TJIS IS T TJ6 ?69?TALB6TJIS IS T TJ6 ?69?TALB6

?6S=LT?6S=LT

@ou must multply by the dluton@ou must multply by the dlutonfactor of 1("factor of 1("

1( 0 ,( + ,(( mg/dL1( 0 ,( + ,(( mg/dL

ser serialdilutions

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