HEMA WEEK 7: CBC AND OTHER ROUTINE PROCEDURES PART 1

Electronic Impedance

Based on the principle that cells produce an electrical impulse whenever suspended in an isotonic solution.

Flow Cytometry

Uses a laser to check for the scattering of light.

Lobularity, granularity, complexity

Flow cytometry check for ______________?

Complete WBC count, WBC differential, RBC parameters, platelets

CBC report includes what?

0.1 mm

Depth of the counting chamber from the coverslip.

5 Large Squares, 1 for RBC and 4 for WBC

Number of significant large squares.

1 mm²

Area of large squares

16 squares, 0.0625 mm²

WBC intermediate square number and size.

25 squares, 0.04 mm²

RBC intermediate square number and size.

400 squares, 0.0025 mm²

Tiny squares number and size.

1:200

Normal RBC count blood to dilution ratio

CBC

Most common test performed in the hematology section

Malignancy and general infection

CBC is a screening test that scans for what?

False

True or False: CBC is able to tell a specific type of blood disease.

Concentrated or Suspended

Automated counting may be done to samples that are?

Hemacytometer

The most commonly used counting chamber in the hematology section.

Red Bead

Part of the Thoma pipette that is used to mix the blood and the diluent.

(101-1)/No. of blood cell counted

Formula for RBC dilution factor

Dacie’s Fluid or Formol Citrate

Best diluent for RBC counting which contains formalin as a preservative

10 mL of 40% Formaldehyde
990 mL of 3% Disodium Citrate

Dacie’s Fluid or Formol Citrate composition

Hayem’s Diluent

Promotes the growth yeasts and clumping of cells. However, it can stand for a long time and has no corrosive effects

Hayem’s Diluent

Mercuric chloride, sodium sulfate anhydrous, crystalline sodium sulfate, water, sodium chloride.

Gower’s solution

Prevents rouleaux formation. Precipitates protein in cases of hemoglobinemia and hyperglobulinemia.

Gower’s solution

Sodium sulfate anhydrous, glacial acetic acid, water.

Toisson’s fluid

High specific gravity and stains WBC. However, supports the growth of fungi.

Toisson’s fluid

NaCl, sodium sulfate, glycerin, methyl violet, water.

Bethell’s fluid

NaCl, sodium sulfate, glycerin, merthiolate (1:1000), water.

Normal Saline Solution (NSS) or Physiologic Salt Solution (PSS)

RBC diluent used in cases of emergency. Useful in the presence of rouleaux formation, and autoagglutination of cell.

Normal Saline Solution (NSS) or Physiologic Salt Solution (PSS)

RBC diluent that is stable and serves as a preservative.

Normal Saline Solution (NSS) or Physiologic Salt Solution (PSS)

Contains sodium chloride and water.

3.8% Sodium Cirtrate

Contains 3.8 g of Sodium Citrate

True

True or False: RBC diluent should be isotonic.

Charging the counting chamber

Most challenging part of cell count

HPO

Objective used in counting RBC

RBC = (total RBC counted) x 10,000

Formula used for standard dilution in RBC counting

RBC = (total RBC counted) x (area counted) x (10) x (dilution conversion factor)

RBC count formula

Recharge the chamber

Supposed a MT counted 165 RBC for the upper chamber and 184 for the lower chamber. What should their next step be?

10^6

Conversion factor from conventional units to SI unit.

3.9-5.5 × 10^12 per L

Reference erythrocyte count for newborns

4.0-6.6 × 10^12 per L

Reference erythrocyte count for 1-3 days old

3.7-5.3 × 10^12 per L

Reference erythrocyte count for 0.5 to 2 years old

3.9-5.3 × 10^12 per L

Reference erythrocyte count for 2 to 6 years old

4.0-5.2 × 10^12 per L

Reference erythrocyte count for 6 to 12 years old

4.5-5.3 × 10^12 per L

Reference erythrocyte count for 12 to 18 years old males

4.1-5.1 × 10^12 per L

Reference erythrocyte count for 12 to 18 years old females

4.5-5.9 × 10^12 per L

Reference erythrocyte count for 18-49 years old males

4.0-5.2 × 10^12 per L

Reference erythrocyte count for 18-49 years old females

Dehydration, exercise, excitement, newborns, high altitude

Physiologic variations resulting to increased RBC count (5)

Testosterone

Males have a higher RBC count due to what?

Polycythemia, Tuberculosis, Fibrosis, Poisoning

Pathologic variations that increase RBC count (4)

Anemia, Blood Loss, Hemorrhage

Pathologic variations that decrease RBC count (3)

Hypotonic and Staining

Diluent for WBC count should be:

11

Maximum capacity of WBC Thoma pipette

Acetic Acid and HCl with methyl or crystal violet

Common diluents of WBC count

Hemolyzing of mature RBC

WBC dilution facilitates what?

Dispersion of RBC

RBC dilution facilitates what?

1:20

Standard dilution ratio of WBC count

1-2 minutes

How long should the cells be able to settle in WBC counting?

LPO

Objective used in scanning and locating the ruled area in WBC counting

WBC = (total cells counted) x 50

Shortcut method for WBC counting in a standard dilution of 1:20

WBC = (total cells counted) x (area) x (10) x (dilution correction factor)

WBC count formula

Recharge, overcharging decreases cell count.

Supposed a MT left the hemacytometer to settle for 10 minutes. What should be the next step?

Count the cells.

Supposed a MT counted a total of 46 in the upper chamber and 56 in the lower chamber for WBC. What should be the next step.

1:20 in WBC Thoma

Supposed the anticipated WBC count is 3.1 to 30. What should be the dilution ratio and thoma pipette used?

1:10 in WBC Thoma

Supposed the anticipated WBC count is 0.1 to 3.0. What should be the dilution ratio and thoma pipette used?

1:100 in RBC thoma with blood until 1 mark

Supposed the anticipated WBC count is more than 30. What should be the dilution ratio and thoma pipette used?

1:200 in RBC thoma with blood until 0.5 mark

Supposed the anticipated WBC count is more than 100. What should be the dilution ratio and thoma pipette used?

DF = (11-1)/Amount of Blood Collected

Diluting factor formula for WBC

4.5 to 11 × 10^9 per L

WBC reference value for adults

10 to 25 × 10^9 per L

WBC reference value for infants

8 to 15 × 10^9 per L

WBC reference value for 1 year olds

Increased WBC

Supposed a patient just had a meal before the collection of blood. How would this affect WBC values.

Increased WBC

Exercise, physical activity, and emotional activity affects WBC in what way?

Lower in the morning, higher in the afternoon

Hourly rhythm of WBC

Correct WBC by using the formula:
(Incorrect WBC [L] x 100)/(No. of RBC + 100)

Supposed a MT counted a total of 7 nucleated RBCs during a WBC count. What should their next step be?

Stained smear is examined microscopically to determine the estimated leukocyte count and to classify leukocytes

WBC diff count principle

EDTA Whole Blood

Specimen requirement for WBC diff count

Methanol

Fixative for WBC diff count

Eosin

Primary stain for WBC diff count

Methylene Blue

Secondary stain

Schilling Counter

Tool used to count each type of WBC

Romanowsky Stain

Commonly used stain for WBC diff count

Wright’s Stain

Routinely used stain in the hematology section

Giemsa Stain

Commonly used for searching for malarial parasites

LPO

Objective used to check for blood film adequacy in WBC diff count

Battlement Track Method

Method used in classifying 100 leukocytes in WBC diff count

Clumped Distribution of Cells

Supposed a MT used the feathered part of a smear for WBC diff count. What will they see?

Crowded and Overlapped Cells

Supposed a MT used either the body or the head part of a smear for WBC diff count. What will they see?

Between the body and the tail using the battlement track method.

Where and how should WBCs be differentiated in a blood film?

Size, N-C Ratio, Cytoplasmic Traits, Nuclear Traits

Criteria for WBC identification (4)

Color and Granules

What should be the focus in checking for cytoplasmic traits of WBCs?

Shape, Chromatin, Presence or Absence of

What should be the focus in checking for nuclear traits of WBCs?

Segmented Neutrophils

Identify the WBC: presence of segmentation, 3-5 lobules

Band Neutrophils

Identify the WBC: no clear segmentation, filament is not pronounced (thicker)

Metamyelocyte

Identify the WBC

Band Cell

Identify the WBC

Band Cell

Identify the WBC

Segmented Neutrophil

Identify the WBC

Segmented Neutrophil

Identify the WBC

Lymphocyte

Identify the WBC: high NC ratio, minimal granules