routine hematology examination

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Routine hematology

examinationRahajuningsih D Setiabudy

Department of Clinical PathologyFaculty of Medicine

University of ndonesia


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Content

De!nition

ndication

Specimen Method

"#ect of storage

nterpretation


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Routine hematology tests

$efore the era of automatic blood cellcounter routine hematology tests consist of

%emoglobin level

&eu'ocyte count

Di#erential count

"rythrocyte sedimentation rate ("SR)

*o+ complete blood count (C$C) consist

of %b, %t, R$C, MC-, MC%,MC%C, .$C, di#/

count, Platelet 0 "SR (manual)


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ndication

1he purpose of hematology examination 21o detect abnormalities of R$C (anemia,

polisitemia)

1o detect abnormalities of .$C (leu'ocytosis,

leu'openia)1o detect abnormalities of platelet

(thrombocytopenia, thrombocytosis)

1o detect abnormalities in leu'ocyte di#erential

count1o detect abnormality of "SR (in3ammation)


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Specimen

$lood is mixed +ith ethylene diaminetetra acetic acid 4 "D15 blood(67"D15 or 68"D15)

Purple top vacuum tube

-enous blood or capillary blood (infantfrom heel pric')

"SR 2 9 volume of blood are mixed +ith: volume of sodium citrate (:/;< M)


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Measurement of%emoglobin

:/ Sahli 4 acid hematin method

less accurate, color develops slo+ly,unstable

8/ %aemiglobincyanide (%iC*) nternationally recommended method

stable and reliable reference preparation

available, 6C* potential ha=ard > sodium

a=ide or sodium lauryl sulphate

7/ ?xyhemoglobin

Standard solution unstable


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%aematocrit or PC-

Microhematocrit method2 accuracyand precision ade@uate for clinicalutility

68"D15 is recommended because67"D15 > shrin'ing of R$C >reducing PC- by 8A

"xcess of anticoagulant > cellshrin'age > falsely lo+ PC-

PC- gradually +ith storage


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$lood cell counting

Manual method2

Diluting the blood +ith speci!cdiluent, (1ur' solution for .$C)

Fill the counting chamber(mproved *eubauer)

Under the microscope count thecell in a certain area (volume)

5utomatic blood cell counter


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5utomatic blood cellcounter

Principles 2 mpedance

blood cells are poor conductors of electricity

+hile certain diluents are good conductors

&ightBscattering technology

Cells pass in single !le in front of a lightsource/

&ight is scattered by the cells passing

through light beam/ Scattered light is detectedby photodiode/ 1he amount of light scatteredis proportional to the volume of cell


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mpedance method

vacuum

blood cells

resistance

direct current(approx/ :;; -)

internal electrodeexternal electrode

ori!ce

sample bea'er

sample

externalelectrode

internal

electrode

ori!ce

vacuum


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Light Scattering system


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"rythrocyte indices

Mean Cell -olume n automated system, MC- is measured

directly

n manual system MC- (PC- x :;)2 R$C Mean Cell %emoglobin

MC% (%b x :;)2 R$C

Mean Cell %emoglobin Concentration MC%C (%b x :;;) 2 PC-

R$C indices are needed to classifyanemia based on morphology


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5utomatic blood cellcounter

Advantages

C- manual

More rapid

&ess volume ofblood is needed

More eEcient

(multiparameter,less humanresources)

Disadvantages

Machine andreagent are

expensive Cumbersome

maintenance

naccurate in someconditions


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Precision of cell counting

Parameter CV ofmanual

method

CV ofautomatic

methodR$C ::A 7A

.$C :A 9A

Platelet 88A G/HA

Wintrobe Clinical Hematology 1974


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5gglutination 2 R$C MC- , MC% MC%C

Microcytosis or many fragmented red cells2R$C , Platelet

%emolysis2 R$C and PC- &ipemic blood2 %b , MC% , MC%C

1+o population of R$C2 MC- normal, RD.

Iiant platelet, clumping, satellitism2 Plt *ucleated red blood cell2 .$C

Sources of inaccuracy inautomatic cell counting


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"rythrocyte sedimentationrate

1he rate of sin'ing R$C is in3uenced by21he di#erence of speci!c gravity

bet+een R$C and plasma

1he capacity of R$C to form rouleaux1he ratio of R$C to plasma

1he plasma viscosity

1he vertically of sedimentation tube1he bore of the tube

Dilution


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"rythrocyte sedimentationrate

"SR is not speci!c test

5ccelerated "SR can be found in 2

5cute and chronic infection due toacuteBphase protein (tbc,rheumatoid arthritis, 5M)

Multiple myeloma due toimmunoglobulin

5nemia


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&o+ "SR

"SR is lo+ (;B: mm) can be foundin 2

Polycythemia

%ypo!brinogenemia

5bnormalities of red cells > diEcult

to form rouleaux 2 poi'ilocytosis,spherocytosis, sic'le cell


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"#ect of storage on theblood count

R$C, .$C, platelet count , and red cellindices stable for J hours/ 5lthough R$Cstart to s+ell

> %t and MC- start 5t 9 C the e#ect on blood count insigni!cant

for up to 89 hours/

t is best to count .$C and especially platelet

+ithin 8 hours

Reticulocyte count begin +ithin hours atroom temp/


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Stability of blood count

Parameter Roomtemperature

Refrigerator

.$C, R$C,Platelet

J hours up to 89hours

Reticulocyte hours 89 hours

%emoglobinconcentration

Up to 8days

days

Dacie and Lewis Practical Haematology !!"


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"#ect of storage onblood cell morphology

Stri'ing changes after :8 K :J hours at roomtemperature

*ucleus stain more homogenously

*uclear lobe > separated,

-acuoles appear in cytoplasm

Cytoplasmic margin > ragged

L hours 2 R$C > crenation and sphering Importance of making blood lm as soon

as possible after blood collection


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*1"RPR"151?* ?F

%"M51?&?IC 1"S1


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%ematological values for normaladults

RC menRC !omen "#$ $#" million%u&'#( $#" million%u&

%b men%b +omen

:H 8 g4d&

:7/H :/H g4d&

%t men%t +omen 45 H A9: H A

MC- men and +omen


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%emoglobin level

%b anemia Red cell indices are needed to classify

anemia2 microcytic hypochromic,

normocytoc normochromic,macrocytic

%b polycythemia 2

Polycythemia veraSecondary polycythemia

Relative polycythemia


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Platelet count

Platelet count 2 thrombocytopeniadecreased production

increased destruction

increased consumptiondilution

pooling

Platelet count

2 thrombocytosis Reactive4secondary

Primary

norma es n eren a


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norma es n eren acount

*eutrophilia 2 bacterial infection,

in3ammation, intoxication, acute bleeding *eutropenia 2 viral infection, typhoid,

chemicals and drug (ben=ene, antithyroid )

&ymphocytosis 2 chronic infection,pertussis, tbc, C&&, mumps

Monocytosis2 tbc, lymphoma, malignancy

"osinophilia2 allergic disorder, parasitic

infection $asophilia2 CM&, P-, ulcerative colitis


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1han' you

routine hematology examination

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