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CBC with Slide Review

An interpretation of analyzer results and the microscopic examination

MELISSA BRAVERMAN, CVT

VETERINARY DIAGNOSTIC LABORATORY

VETERINARY SPECIALTY CENTER

COMPLETE CBC•Analyzer results

•Slide review�Platelets

�WBC differential

�WBC morphology

�RBC morphology

ANALYZERS

• Human error

• Large variation in animal blood cells

compared to humans

• Feline platelets can be normally 2x

the size of other mammals platelets

and therefore are often mistaken for

RBCs

ANALYZER RBC INDICESHGB – Hemoglobin

HCT – Hematocrit

MCV – Mean Corpuscular (Cell) Volume

MCH – Mean Corpuscular (Cell) Hemoglobin

MCHC – Mean Corpuscular (Cell) Hemoglobin Concentration

RDW – Red Cell Distribution Width

HEMOGLOBIN (HGB) g/dLDirect measurement of blood’s oxygen carrying capacity

Can be falsely increased by hemolysis, lipemia and Heinz bodies

Directly measured by instrument

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HCT/PCV %

HCT on analyzer is a calculation

(MCV x RBC) / 10

• Use H:H to check for accuracy of HCT. Hemoglobin: Hematocrit ratio

Hgb x 3 = Hct (within a margin of 4%)

Hgb = 16.2 16.2 x 3 = 48.6 HCT = 46.8%

• INCREASED H:H RATIO GENERALLY CAUSED BY HEMOLYSIS OR LIPEMIA

• LIPEMIA AND HEMOLYSIS MAY FALSELY INCREASE Hgb AND RBC INDICIES

• RECALCULATE MCHC USING SPUN PCV

MCV (fL)

Mean Corpuscular Volume – Average size of single RBC

MCV is directly measured by analyzer.

MCV can be determined indirectly by calculation but it tends to be less accurate.

(HCT x 10) / RBC = MCV

(46.8 x 10) / 6.63 = 70.6

Increased MCV – Macrocytosis

Decreased MCV - Microcytosis

MCH (pg)

Mean Corpuscular Hemoglobin – Average amount of Hgb inside average RBC

(Hgb x 10) / RBC = MCH

MCH provides no added value because the interpretation of the anemia depends on the MCV and MCHC

MCHC (g/dL)

Mean corpuscular hemoglobin concentration – Average Hgbconcentration within RBCs

(Hgb / Hct) x 100 = MCHC

(16.2 / 46.8) x 100 = 34.6

Increased MCHC values are artifact generally caused by lipemia, hemolysis and presence of Heinz bodies.

•Normal MCHC – Normochromic

•Decreased MCHC - Hypochromic

MORPHOLOGIC ANEMIA CLASSIFICATION

•Normocytic normochromic

� Chronic renal disease

•Macrocytic hypochromic

� Regenerative anemias with marked reticulocytosis

•Macrocytic normochromic

� Regenerative anemias

� FeLV

•Microcytic normochromic/hypochromic

� Chronic iron deficiency

� Portosystemic shunt

RDW %

RBC distribution width – amount of variation in size of RBCs

Anisocytosis (Size)

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REFERENCE RANGES (from University of Illinois Veterinary Diagnostic Laboratory)

TEST UNITS CANINE FELINE

HCT % 35 – 52 30 – 45

HEMOGLOBIN g/dL 12 – 18 8 – 15

RBC COUNT x106 5.5 – 8.5 5 – 10

MCV fL 60 – 77 39 – 55

MCH pg 20 – 25 13 – 18

MCHC g/dL 32 – 36 30 – 36

WBC COUNT x103 6.0 – 17.0 5.5 – 19.5

NEUTROPHIL/SEG x103 3.0 – 11.5 2.5 – 12.5

BANDS x103 0.0 – 0.3 0.0 – 0.3

LYMPHOCYTE x103 1.0 – 4.8 1.7 – 7.0

MONOCYTE x103 0.2 – 1.4 0.0 – 0.9

EOSINOPHIL x103 0.1 – 1.0 0.0 – 0.8

BASOPHIL x103 0.0 – 0.2 0.0 – 0.2

PLATELET COUNT x103 200 - 900 300 - 700

SLIDE REVIEW

•Wipe slide clean

•Let blood drop spread across spreader slide

•Do not lift spreader slide

•If HCT is high, decrease angle of spreader slide

•If HCT is low, increase angle of spreader slide

SLIDE REVIEW*ALWAYS SCAN FEATHERED EDGE FIRSTALWAYS SCAN FEATHERED EDGE FIRSTALWAYS SCAN FEATHERED EDGE FIRSTALWAYS SCAN FEATHERED EDGE FIRST

SLIDE REVIEWDIFFERENTIAL

CANINE FELINE

SLIDE REVIEWPLATELETS

NORMAL FELINE PLATELETSNORMAL CANINE PLATELETS

SLIDE REVIEWPLATELETS

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SLIDE REVIEWPLATELETS

Canine

Platelet count = 0.0

SLIDE REVIEWPLATELETS

CALCULATING PLATELET ESTIMATE

COUNT AMOUNT OF PLATELETS SEEN ON 10 VIEWS USING 100X OIF

TAKE AVERAGE OF COUNTED PLATELETS AND MULTIPLY BY 15,000 – 20,000

Ex. – COUNTED 104 PLATELETS TOTAL ON 10 VIEWS

104/10 = 10.4 10.4 X 20,000 = 208,000 (208k)

NOTE IF CLUMPS SEEN (RARE, FEW, MODERATE, MANY)

NOTE IF LARGE PLATELETS SEEN (RARE, FEW, MODERATE, MANY)

WBC COUNTABSOLUTE VS. RELATIVE

WBC DIFFERENTIAL RELATIVE VALUES ARE THE % OF EACH TYPE OF WBC COUNTED

WBC DIFFERENTIAL ABSOLUTE VALUE IS THE % MULTIPLIED BY THE WBC COUNT

CANINE WBC COUNT = 10.1 K/ul

RELATIVE VALUE ABSOLUTE VALUE

NEUTROPHIL 60% 0.6 X 10.1 = 6.06 K/uL

LYMPHOCYTE 24% 0.24 X 10.1 = 2.42 K/uL

MONOCYTE 10% 0.1 X 10.1 = 1.01 K/uL

EOSINOPHIL 6% 0.06 X 10.1 = 0.61 K/uL

BASOPHIL 0%

WBC COUNTABSOLUTE VS. RELATIVE

WBC COUNT = 2.5 K/ul

RELATIVE VALUE ABSOLUTE VALUE

NEUTROPHIL 15% 0.15 X 2.5 = 300 uL

LYMPHOCYTE 80% 0.8 X 2.5 = 1,600 ul

•NEUTROPENIA

•RELATIVE LYMPHOCYTOSIS DUE TO NEUTROPENIA

•NOT ABSOLUTE LYMPHOCYTOSIS

WBCNEUTROPHILS

WBCHYPERSEGMENTED NEUTROPHILS

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WBCTOXIC NEUTROPHILS

WBCBAND NEUTROPHIL

WBCBAND NEUTROPHILS

•A left shift is an increase in number of immature neutrophils

•Most common cause of a left shift is inflammation

•Regenerative left shift – mature neutrophils outnumber bands

•Degenerative left shift – bands/immature neutrophils outnumber mature

WBCLYMPHOCYTES

WBCLYMPHOCYTES

Reactive lymphocyte – cytoplasmic basophilia

Atypical lymphocyte – increased cytoplasmic volume

WBCLYMPHOBLAST

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WBCMONOCYTES

WBC EOSINOPHILS

Canine Feline Degranulated

WBCBASOPHILS

CANINE FELINE

MAST CELLS

RBC Normal FelineNormal Canine

RBCPolychromatophil & nRBC

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RBC

POLYCHROMATOPHILS RETICULOCYTES

RBCHeinz Bodies

RBCHeinz Bodies

New Methylene Blue

RBC

Echinocyte Echinocyte

RBC

Acanthocyte Keratocyte

RBC

Target Cell Ovalocyte

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RBCHypochromic Cell Torocyte

RBC

Spherocytes Howell Jolly Body

RBC

Rouleaux Agglutination

SMUDGED RBCs

RBCSCHISTOCYTE

POLYCHROMATOPHILS

1+ 2+ 3+ 4+

Canine 2-7 8-14 15-29 >30

Feline 1-2 3-8 9-15 >15

Bovine/Porcine 2-5 6-10 11-20 >20

Equine Rarely

observed

All other morphology

Rare Few 1+ 25% 2+ 50% 3+ 75% 4+ 100%

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CASE REVIEW13 yo F/S Persian presented for anorexia and lethargy

mm = pale pink, depressed; other vitals= WNL

CBC on analyzer

WBC – 13.1 RBC – 7.06 M/Ul

Neu – 80% HGB – 11.2 g/dL

Lymph – 15% HCT – 33.6%

Mono – 3% MCV – 47.6 fL

Eos – 0% MCHC – 33.4 g/dL

Baso – 2% RDW – 19.3%

Platelets – 32.0

CASE REVIEW

CASE REVIEW CASE REVIEW

CASE REVIEWDifferential

Neut – 27%

Lymph – 6%

Mono – 0%

Eos – 1%

Baso – 0%

Blasts – 66%

Platelet est – 120K /uL

Rare giant platelet

Pathologist review - The blasts have eccentrically-placed, primarily round nuclei with stippled chromatin and prominent nucleoli, with a small rim of deeply basophilic cytoplasm that rarely contains few small discrete vacuoles. These blasts appear lymphoid (which can be confirmed with flow cytometry or ICC on unstained blood smears). Differentials include acute leukemia (likely lymphoid, less likely erythroid), and stage 5 lymphoma (if other tissues are involved). AU 11-14-16

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