Unit 3.5 DIFFERENTIAL COUNT Flashcards by Arriane Cyrel (MTI)

This involves both quantitative and qualitative study of the types of leukocytes.

DIFFERENTIAL COUNT

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It also includes quantitative and qualitative study of thrombocytes as well as morphologic abnormalities of erythrocytes

DIFFERENTIAL COUNT

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represents the percentage distribution of the different WBCs

DIFFERENTIAL COUNT

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METHODS OF BLOOD SMEAR PREPARATION

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Preferred for bone marrow preparations

Coverslip Method

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It has the advantage of superior leukocyte distribution

Coverslip Method

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NOT for routine peripheral blood smear preparation.

Coverslip Method

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Coverslip Method Procedure:
Place (?) at the center of coverslip held between thumb & forefinger
Invert another coverslip rotated (?)
As soon as the blood spreads, pull apart the 2 coverslips in a (?).

1 drop of blood
45O
firm and steady motion

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Glass Slide-Coverslip Method

Place 1 drop of blood near (?) of a glass slide. Place coverslip.
As soon as the blood spreads, (?) the coverslip along the surface of the slide

one end
push forward

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Most convenient and most commonly used.

Two-Glass Slide Method (Wedge slide/Push smear)

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Two-Glass Slide Method (Wedge slide/Push smear) Disadvantages:
1. Tendency of larger cells to settle at the slide edges & (?)
2. (?) of nucleated cells
3. (?) to cells

feathered ends

Poor distribution

Greater trauma

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Utilizes the wedge procedure

Miniprep/Hemaprep

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A precision blood spreader prepares dual smears simultaneously at a constant angle and speed.

Miniprep/Hemaprep

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Glass slides are placed on a platform inside the instrument

Hemaspinner

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Instrument spins causing the slide to be covered with a monolayer of cells beam of sensor light to detect blood distribution

Hemaspinner

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For finding reactive/immature or abnormal cells that are present in small numbers

Buffy Coat Smear

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For easier location of bacteria & parasites

Buffy Coat Smear

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Buffy coat smear is considered a concentrate smear for WBCs, thus it recommended when the WBC count is _________.

Buffy Coat Smear

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For blood parasites examination.

Thick Blood Smear

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CHARACTERISTICS OF A PROPERLY PREPARED WEDGE SMEAR
1. Length: must occupy at least (?) of the slide
2. Should be free of (?)
3. Should have (?) appearance
4. Should be (?) than the stationary slide
5. Must have (?) from thick to thin
6. (?) to allow proper fixation
7. Should terminate in a (?)
8. Should contain at least (?) in which 50% of the RBCs do no overlap each other; the remainder ma overlap in doublets or triplets and the central pallor should be clear

1/2 to 2/3

waves, holes, and ridges

smooth and even

narrower

gradual transition

Thin enough

feathery tail

10 LPF

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Too large drop of blood

TOO THICK SMEAR

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Too fast spread

TOO THICK SMEAR

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Too high angle

TOO THICK SMEAR

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High hematocrit

TOO THICK SMEAR

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Too small drop of blood

EXTREMELY THIN SMEAR

Too slow spread

EXTREMELY THIN SMEAR

Too low angle

EXTREMELY THIN SMEAR

Low Hematocrit

EXTREMELY THIN SMEAR

Accumulation of nucleated cells

GRITTY APPEARANCE

large number of leukocytes

GRITTY APPEARANCE

Use of heparin as anticoagulant

GRITTY APPEARANCE

Too slow spreading/delay in spreading

GRITTY APPEARANCE

Using only a part of the drop of blood

GRITTY APPEARANCE

Rough edge or dirty spreader slide

GRITTY APPEARANCE

METHODS OF MANUAL DIFFERENTIAL COUNT

WBCs are counted in consecutive fields as the blood film is moved from side to side

Cross-sectional/Crenellation Method

WBCs are counted in consecutive fields from the tail toward the head of the smear

Longitudinal Method

Ideal if smear is thin enough since cells can be identified all the way.

Longitudinal Method

WBCs are counted in a pattern of consecutive fields near the tail on a horizontal edge. Count 3 consecutive horiontal edge fields, 2 fields towards the center, 2 horiontal and 2 vertical to the edge

Battlement Method

Using the Oil Immersion Objective: Estimate (?); Examine (?) Perform WBC Differential count - A total of (?) are counted and classified according to type

Platelets and WBCs RBC morphology 100 WBCs

The total number of cells to be counted could be decreased or increased when any of the following criteria are observed: (?) if the total WBC count is < 1 x 10 9/L (< 1,000/µL) Count (?) if in the first 100 count, any of the following are observed::

50 Cells additional 100 cells

>(?) Eosinophils > (?) Basophils >(?) Monocytes (?) are greater than Neutrophils

10% 2% 11% Lymphocytes

METHODS OF CLASSIFYING NEUTROPHILS

WBCs are classified based on the number of lobes indication their age

ARNETHS CLASSIFICATION

- WBCs are classified according to granulation

SCHILLINGSS CLASSIFICATION

Younger forms are placed on the left; mature and old forms on the right

SCHILLINGSS CLASSIFICATION

1 round or indented nucleus

5% (blast cells)

2 nuclear divisions

35 %

3 nuclear divisions

III

41 %

III

4 nuclear divisions

17 %

5 or more

2% (oldest)

Schillings Hemogram

1 2 3 4 5 Eos Baso Lym Mono

Schillings Hemogram Shift to the LEFT:

increase in immature forms

Schillings Hemogram Shift to the RIGHT:

- increase in mature forms

young forms with accompanying WBC count.

Regenerative Shift to the Left

Infections, appendicitis & acute sepsis

Regenerative Shift to the Left

young forms with N or WBC count

Degenerative shift to the Left

Typhoid fever and TB

Degenerative shift to the Left

Pernicious anemia or megaloblastic anemia

Shift to the RIGHT

: cells that possess segments or lobes connected with a distinct thin chromatin filament

Filamentous

: nucleus with thinned-out portion

Non-filamentous

REPORTING 1. Relative count - Cells are reported in 2. Absolute count - Cells are reported as

% or decimal number per volume of blood (e.g.: cells /µL or /L )

Absolute count =

Relative count (%) x total WBC count

47 77%

PM Neutrophil

1.8 7.8 x 109/L

PM Neutrophil

0 7%

Band cells

0 0.7 x 109/L

Band cells

20 40%

Lymphocytes

1.0 4.8 x 109/L

Lymphocytes

2 10%

Monocytes

0.01 0.8 x 109/L

Monocytes

0 6%

Eosinophils

0 0.6 x 109/L

Eosinophils

0 1%

Basophils

0 0.2 x 109/L

Basophils

are volatile

Circulating cells

cause lymphocytes & eosinophils to disappear from circulation within 4 8 hrs

Corticosteroid hormones

causes granulocytosis within minutes

Epinephrine

CLINICAL SIGNIFICANCE of variation in counts:

Acute Infections -