blood exercises (phyana lab)

7/24/2019 Blood Exercises (PHYANA LAB)

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PHY-ANA LABBLOOD

EXERCISES


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COMPLETE BLOOD COUNT

Hematocrit

Hemoglobin Differential white blood cell

Red blood cell

White blood cell


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HEMATOCRIT

% of formed cells in whole blood

99% RBCs and 1% WBCs and platelets

Estimate if RBCs are adequate

Greek hematoblood andcrit to

judge

Aka packed cell volume (PCV), Hctor erythrocyte volume fraction (EVF)


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HEMATOCRIT

One of the simplest, most accurate, & valuable tests

Detecting cases of anemia


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HEMATOCRIT

Specimen

Fresh capillary blood (with heparin)

Adams Microhematocrit Method


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HEMATOCRIT

1. Blood of thecapillary tube

2. Sealing clay (3 mm)

3. Centrifuge 10,000rpm for 4-5 minutes

4. Level of packed RBCusingmicrohematocritreader


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HEMATOCRIT

Capillary tube withseal towards the

outside

Balance tubes inthe centrifuge

Securely screw thecover of thecentrifuge


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HEMATOCRIT

When rotation has stopped,

remove tube Take note of the appearance


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HEMATOCRIT

Capillary tubewith seal

toward thecenter

Align upper

portion of theseal with theblack line


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HEMATOCRIT

Rotate thewhole assembly

so that the pinstops (100mark)

Rotate theupper disk tomove the curve

line with the top


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HEMATOCRIT

Rotate the entire assemblyuntil the curved line is lined

up with the boundarybetween packed RBC andplasma

Read the % packed cells atthe right


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HEMATOCRIT


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HEMOGLOBIN

Red-pigmented protein

Transports oxygen andcarbon dioxide

Measured as

oxyhemoglobin Indirectly measured byconverting to compounds


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HEMOGLOBIN

Acid Hematin

Method Yellowish brownsolution is comparedto the color standard

in the comparatorblock

Darker the color =higher Hgb content


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HEMOGLOBIN

1. 0.1N HCl2 mark of Sahlis tube

2. Aspirate 0.02 mL blood using Sahlis pipette

3. Expel the blood sample to the tube4. Rinse the pipette with dist. water 3xadd to the

mixtureStand 10 mins

5. Add dist. water drop by drop (mix with stirring rod)

until color matches with the block6. Readinglower meniscus

7. Report gm% or gm/dL or gm/100mL (CU) and ingm/L (SI)


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DIFFERENTIAL WBC COUNT

Examination of a thin smear determining the

percentages of WBC types


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Specimen

EDTA blood

Within 2-3 hours ofcollection

Within the mark of thetube

Avoid

Old specimen

Excessive amount of

anticoagulant to specimen


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Blood smear

preparation Most important step

Two-Slide or Wedge

Method Simplest

Most popular


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1. Drop of blood from mixed

sample on a clean glass slide2. Spreader slide at an angle of

about 30-45o

3. Allow blood to spread evenly;Control thickness of smear

4. Air dry

Do not blow dry: RBC artifact


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Good smear:

Thick to thin Occupy 2/3 or

Smooth and evensurface

Free from ridges,waves, holes

Margin-free

Feathery edge

Factors that affect:

Angle of the spreaderslide

Greater angle: thicker &shorter

Size of the blood drop Speed of spreading


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Staining of Blood Smears

Methanol: fixative (30s) Eosin: acidic dye (6s)

Stains Hgb & leukocytes

Methylene blue: basic dye

(4s) Nucleoproteins, nucleic acids

Buffer solution (pH 7.2) for

45s


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Staining of BloodSmears

Dip Method Rapid)

Quick method

Modified Wright-Giemsa

buffered in methanol atpH 6.8

Tightly sealed


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Blood Smears

RBC: pink to salmon Nucleus: dark blue to purple

Neutrophils: lavender to lilac

Basophils: dark blue to black Eosinophils: red to orange

Area between cells: colorless,clean and free of precipitates


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Smear Examination

1. LPO Assess overall quality Rapid detection of large

abnormal cells

Not overlapping or too scanty2. Shift to OIO


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Method of Differential Counting

Battlement Count 100 white blood cells whiledifferentiating


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Method of Differential Counting

Battlement Count 100 white blood cells whiledifferentiating


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HEMACYTOMETER

Counting chamber

WBC pipette RBC pipette

Accessory devices

Suction device Thick cover slip


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COUNTING CHAMBER

Heavy, colorless glass

3 parallel platformsseparated by moats

Central: 0.1 mm lowerthan lateral

Transverse groove


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COUNTING CHAMBER


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COUNTING CHAMBER

1 Primary square

3x3 mm (9 sq. mm)

9 Secondarysquares

1x1 mm

4 corners: WBCcount

16 tertiary squares

W1, W2, W3, W4 64 squares


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COUNTING CHAMBER

Central secondarysquare

25 tertiary squares 0.2 mm each

16 quaternarysquares

Total number ofquaternary: 400

RBC count

5 tertiary squares:80 squares


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DILUTED BLOOD

PREPARATION 0.5 mark: blood

Diluting fluid

11 WBC, 101RBC

Constant

rotation Over aspirate

or presence of

bubbles: repeat


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CHARGING

Cover slip

No dirt, thumb marks,

tissue strands Discard

WBC 2-3 drops

RBC 5-6 drops Angle of the pipette

(30-35)

Stand for 5-10 minutes


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RBC AND WBC THOMA

PIPETTES RBC pipette

Stem 0.0 to 1.0 contains1 unit of volume

Mixing chamber or bulb0.1 to 101 holds 100units of volume

WBC pipette

Stem 0.0 to 1.0

Bulb 1.0 to 11

Stem volume is 10x the

bulb volume: 10 units

RBC

PIPETTE

WBC

PIPETTEUpper mark 101 11

Bore Smaller Bigger

Bead Red White

Dilution 1:200 1:20

Size of bulb Bigger Smaller


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STANDARD PATTERN OF

COUNTING Cells touching

any of the lines

on the top andleft borders areincluded

Cell difference

between 2squares

RBC 20 or less

WBC 12 or less


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COUNTING CELLS


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COMPUTATION

RBC COUNT No. of cells/cumm = total number of cells counted

area X depth X dilution

= total number of cells counted

1/5 X 1/10 X 1/200

= cells counted X 10,000

Normal values: male: 4.5-6.0 M/cumm

female: 4.0-5.5 M/cumm


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COMPUTATION

WBC COUNT

No. of cells/cumm = total number of cells counted

area X depth X dilution

= total number of cells counted

4 X 1/10 X 1/20

= cells counted X 50

Normal value: 5,000-10,000/cumm


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BLOOD GROUPS

Antigens

Antibodies

Agglutination


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ABO BLOOD GROUPING


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BLEEDING TIME

ukes Method

Finger prick

Allow blood to flow freely

Start time: drop of bloodappears

Blot with filter paper Do not touch the wound

Stop time: when bleeding stops

Normal: 1-3 minutes


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COAGULATION TIME

Drop or Slide Method

Prick

Drop of blood on a slide

Start: when in contact with the slide

Tip of lancet every 30 second

interval Observe fibrin formation

Stop timer

Normal: 3-6 minutes


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COAGULATION TIME


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HYPEREMIA OR

CONGESTION Note the skin color, blood vessel

condition, temperature of left indexfinger

Immerse in hot water (60C) for 5minutes

Note the changes and thesensation felt

Rubber band (5 minutes)2ndinterphalangeal joint

Note the changes and thesensation felt


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HYPEREMIA OR

CONGESTION Hyperemia: active increase in

blood volume

Dilation

Physiological: blushing or duringexercise

Reddish

Congestion: passive increase in

volume of blood Impaired venous blood flow orvenous obstruction

Reddish-blue (cyanosis)

Always pathological

Cardiac failure


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CAPILLARY RESISTANCE

TEST Assesses the fragility

of capillary walls

Hemorrhagictendency

Thrombocytopenicpurpura


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TEST Thrombotic Thrombocytopenic Purpura

(clots)-(low platelet number)-(purple bruises)

Rare blood disorder

Blood clots form in capillaries

Uses up platelets Bleeding problems


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TEST Tourniquet Test Rumpel-Leede or Hess)

Mark red spots on the arm

Wrap the cuff of sphygmomanometer around

Inflate to 100 mmHg (5 mins) or 50 mmHg (10mins)

Release pressure (15-20 mins elapse)

Count the number of petechiae (ventral)

Interpret results


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TEST Interpretation of results

Number of petechiae Grade

0-10 1+

11-20 2+

21-50 3+

51 and above 4+

blood exercises (phyana lab)

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