Hematology & Body Fluids Lab Practical - Study Guide

Hematocrit

Male: 42 - 52%

Female: 36-46%

Hemoglobin

Male 14.0 - 17.4 g/dL

Female: 12.0-16.0 g/dL

RBC Count

4.5 - 5.5 × 10^6 uL / 10^12 L

WBC Count

4.5 - 11.0 × 10³ uL / 10^9 L

What are the RBC indices and their significance?

• MCV (Mean Corpuscular Volume):

  • avg vol. of RBC → diagnose types of anemia.

• MCH (Mean Corpuscular Hemoglobin):

  • avg amt of HgB in a single RBC; indicates oxygen-carrying capacity.

• MCHC (Mean Corpuscular Hemoglobin Concentration):

  • conc. of HgB in RBCs → identify types of anemia.

• RDW (Red Cell Distribution Width):

  • Measures the variation in size of RBCs → cause of anemia.

MCV Value for Adults

80 - 100 fL

MCH Value for Adults

28 - 34 pg

MCHC Value for Adults

32 - 36%

RDW Value for Adults

12.0 - 14.6%

WBC Differential Values

• PMS: 40-80%

• Bands: 0-5%

• Lymphs: 25-35%

• Monos: 2-10%

• Eos: 0-5%

• Basos: 0-1%

Absolute Values for WBC Diff

• PMN’s/Bands: 1.8 - 7.0 × 10^9 / L

• Lymphs: 1.0 - 4.0 × 10^9 / L

• Monos/Eos/Basos: 0.2 - 1.0 × 10^9 L

Plt Count

150,000 - 450,000 uL or 150 - 450 10^6 L

Spun Hematocrit (manual method)

34-50%

Reticulocyte Count

0.5 - 2.0%

Rule of 3 for RBC indices calculations

1. Hgb x 3 = HCT

2. RBC x 3 = Hgb

3. RBC x 9 = Hct

What is the principle of the reticulocyte count procedure?

immature RBCs with RNA and organelles that can be stained with vital dyes, forming a filamentous network. They appear polychromatophilic on Wright’s stain (Methylene Blue Reagent) and assess bone marrow erythropoietic activity.

Reticulocyte Count Procedure

1. 2 drops of methylene blue + 2 drops of well-mixed blood

2. Mix & incubate for 15 mins

3. 2 smears + Air Dry

4. 100x until 500+ total cells counted for each slide

5. (Divide sum of reticulocytes by total number of RBCs)100

• Results should be within 10% of each other

Reticulocyte Count Formula

% Retics = (# of retics x 100)/1000 RBCs

Platelet Clumps Procedure

• Check specimen for clot

• Make peripheral smear and stain

• Under 50x objective lens, check for plt clumps, fibrin strands, satellitosis → WBC estimate

  • 100x objective to perform a plt estimate

• Plt. estimate should be ±20% from the analyzer result

Platelet Clumps Significance

Source of interference → causes falsely low plt count

• incubate at 37C for 15-30 min

• Vortex aliquot of sample (only 2 min to not activate plts → cause clumps)

• Draw another EDTA & blue top tube if too many clumps

Corrected WBC Count (due to nRBC)

Correct any total WBC count per mm3 that has greater than 5* nRBCs/100 WBCs counted.

Platelet Estimate Calculation and Ranges

• Decreased: 4-9 plt/field

• Normal/Adequate: 10 – 30 plt/field.

• Increased: 31-45 plt:

• determine the avg number of platelets per field using 5 – 10 different fields (100x) and multiply this result by 15,000.

Serous Fluids Lining Cells are called?

Mesothelial Cells

CSF Lining Cells

• Choroid - Lining of Choroid Plexus

• Ependymal - Lining of Ventricles of neural

• Spindle - Arachnoid Membrane Lining

Choroid Plexus Cell

Synovial Lining Cell

Synoviocytes

• Eccentric nucleus

• “Fried Egg” appearance

• Possible debris inside cytoplasm

Serous Fluid Lining Cell

Bronchoalveolar Lavage Fluid (BAL) Lining Cell

Macrophage and Ciliated Cell

Malignant Cells

• Very large, basophilic cells

• Irregular shaped nuclei and nucleoli

• Uneven staining in the cytoplasm

• Various nuclear size and shape

• Vacuoles over the nucleus

Signet ring vs. LE cell

• Signet cells = Large Vacuole, nucleus flattens to one side → many: possible malignancy

• LE cell (Lupus erythematosus) = ingested WBC within vacuole (pink blob) → autoimmune disease

Macrophage in Body Fluid

Monocyte in Body Fluids

Manual Platelet Count Procedure

• 40x = Within each of the small 25 squares, count cells touching the top & left-hand borders. If the border has 3 parallel lines, use the middle line as the border.

  • Count both sides of the hemocytometer. The difference should be 10% or less. Take the avg of the count from both sides.

• plt count X 1000 = plt/μL

• plt normal range: 150,000 – 450,000/μL or 150 – 450 X 10^9/L

Waiting times for plt count

• 5 mins for cell lysis to complete

• Place the hemocytometer into the humidity chamber (petri dish) and allow the cells to settle for 5-10 mins