Eosinophil Count For normal adrenal function: second eosinophil count is atleast
50% less than the first count (because there is a temporary
1. State the diagnostic importance of Eosinophil Count
- Hypoadrenalism (Addison's): increase
2. State the principle of the test.
• Counting Chamber
1) Neubauer - not recommended for eosiophil count due to small
volume ; uses the same procedure for WBC count except for blood
drawn
Principle: The number of eosinophil can be accurately determined
directly by counting in a hemocytometer. Whole blood is diluted
by a use of a special diluting fluid that stains eosinophils and
hemolyzes RBCs and other leukocytes leaving only eosinophils in
contact.
Procedure:
Vol of blood drawn is up to 1 mark
Allow to stand RT for 20 mins
Count in 9 squares
Should be counted within 30 mins after dilution
(to prevent disintegration)
2) Fuchs-Rosenthal - with 2 counting areas
3) Speirs-Levy - with 4 counting areas
• Thorn's Test / Eosinophil Depression Test
- asses normal adrenal cortical function
- affects both eosiphils and basophils
Principle:
Procedure:
Px is on a fasting state from 8:00 pm
Determine baseline eosinophil count early the next mornkng
Inject 25mg of ACTH
Determine eosiophil count after 4 hrs
Interpretation:
increase of ACTH)
Hyperadrenalism (Cushing's): decrease
3. Identify the reagents and materials used in the procedure.
Whole blood anticoagulated with EDTA or heparin
Phloxine
WBC Pipet
hemocytometer
microscope
4. Explain the procedure of Eosinophil Count
1. Draw well mixed anticoagulated blood to the 1 mark of the
WBC Thoma pipet.
2. Wipe the outside portion of the pipet.
3. Draw the diluting fluid (phloxine) to the 11 mark.
4. mix for 10 minutes using a pipetter shaker
5. expel the first 4 drops and charge to both sides of the counting
chamber
6. allow the cells to settle for at least 3 minutes before counting
7. count the eosinophils in the entire ruled area of both sides of the
hemocytometer (9 large squares)
8. get the average count and compute for the absolute counting
eosinophil/mm³ = (no. of eosinophil counted / 9) x 100
5. Identify the potential sources of errors.
Wrong dilution
Contaminated diluting fluid
Can't recognize eosinophil
Technical errors
6. Identify the conditions that may yield abnormal Eosinophil
Count results.
Parasitic infections
Allergic reaxtions
Leukemia
7. Identify the standard reference value.
150-300/mm³ of blood
D-Dimer Test
1. State the diagnostic importance of D-Dimer Test
2. State the principle of the test.
3. Identify the reagents and materials used in the procedure.
4. Explain the procedure of D-Dimer Test
5. Identify the potential sources of errors.
6. Explain the pathophysiology of DIC.
7. What are the diseases or conditions that can cause DIC?
Lupus Erythematosus Cell Preparation
1. Give the clinical significance of LE Cell.
- to diagnose SLE
Discoid LE – skin lesions with scaling folicular plugging
Systemic LE – facial erythema, rheumatic arthritis,
photosensitivity; molar rash, reynaud’s phenomenon
2. State the principle of the test.
Cells are damaged to allow liberation of nuclei from cells. Nuclear
antibodies present in the serum bind to the liberated nuclear mass
facilitating phagocytosis of neutrophils. Upon incubation at 37C,
neutrophils ingest the liberated nuclei. The whole blood is then
centrifuged and the buffy coat is smeared and microscopically
examined for the presence of neutrophils with phagocytized
homogenous nuclear mass.
3. Explain the procedure on LE cell preparation.
A. Clot Method
1. Collect 10mL venous blood from the patient and allow the
blood to coagulate
2. Macerate the clot and serum with the use of applicator sticks or
let the clot pass through a wire sieve.
3. collect the macerations and transfer to several Wintrobe tubes.
Incubate the tubes for 1 hour in a water bath at 37C.
4. Centrifuge the tubes at 3500 rpm for 30 minutes.
5. Carefully remove the serum and transfer the buffy coat near the
end of each glass slide.
6. Smear, dry, and stain the Wright’s stain.
7. Examine stained smears microscopically for the presence of LE
Cells.
B. Rotary or Defibrinated Method
1. Place 10 mL of freshly collected venous blood into an
erlenmeyer flash with glass beads.
2. rotate the flask in figure of 8 motion for at least 30 minutes.
3. Remove the glass beads.
4. Transfer the rotated and defibrinated blood into several
wintrobe tubes and incubate in a water bath at 37C for 1 hour.
5. Centrifuge the tubes at 3500 rpm for 30 minutes. Carefully
remove the serum and transfer a drop of buffy coat near one end
of each slide and smear.
6. Let the blood smear dry and stain with Wright’s stain.
7. Examine stained smear under the microscope for the presence
of LE Cells.