Download - 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