Group 8 September 23, 2021

Gonzales, Victor Lorenzo Tue, 11:00-2:00 PM

Molino, Charmaine
Ninalga, Karla Khrystina
Oloan, Daisybeth
Paguel, Lanlie Ericka

Questions for Research:

1. Differentiate between Absolute count and relative count (6 points)
 Relative count is a type of WBC differential count that measures each type of leukocytes in
relationship with the total WBC count, and is expressed usually as a percentage or decimal.
Absolute count, on the other hand, is the actual number of each type of leukocytes in the
blood sample. It is not measure directly, but calculated by multiplying the blood cell count
by the relative count in the sample (Absolute count = Relative count in % x total WBC
count). Of these, the absolute value is much more important than the relative value. For
example, a 70% Relative Neutrophil Count may seem within normal limits. However, if the
total WBC is 30,000, the absolute value (70% x 30,000) of 21,000 would be an abnormally
high count.

References:
Glowm. (n.d.). White Blood Cell Differential Count (Differential, Diff). https://glowm.com/
resources/glowm/cd/pages/resources/Lab/white.htm
Sight Dc. (n.d.). Neutrophils: definition, absolute high & low, ranges.
https://www.sightdx.com / knowledge-center/neutrophils

2. Explain why manual RBC count is unreliable. (5 points)

 Manual counting of RBCS is unreliable because there are too many RBCs shown in the
counting chambers of a hemocytometer, and their relatively minute size also adds to the
manual method’s disadvantage. This could lead to miscounting errors such as recounting
the same RBC or failing to count (skipping) an RBC or two. These errors would produce
erroneous results like a falsely increased and falsely decreased counts of RBC, respectively.
Further, manual RBC counting relies on human visualization, judgment, and skill, so it is
even more unreliable and inaccurate results may be produced.

References:
Greer, J. et. al (2014) Wintrobe’s Clinical Hematology 13th edition. pp. 32. Wolters Kluwer I
Lippincott Williams and Wilkins.
Science Direct. (2020). Errors in Red Blood Cell count. https://www.sciencedirect.com/topics
/biochemistry-genetics-and-molecular-biology/erythrocyte-count
3. Why is it necessary to compute for corrected WBC count in the presence of significant
number of nucleated red blood cells? Give the formula (5 points)
 If five or more nucleated RBCs per 100 WBCs are observed on the differential count on a
stained peripheral blood film, the WBC must be corrected for these cells. Because
nucleated RBCs are indistinguishable from WBCs when seen on the hemocytometer, they
could be erroneously counted as such, and the result would show a false increased in
WBC count. This could lead to misdiagnosis and improper treatment of the patient, so it’s
important to compute for correct WBC count. It is accomplished by using the formula:

𝑈𝑛𝑐𝑜𝑟𝑟𝑒𝑐𝑡𝑒𝑑 𝑊𝐵𝐶 𝑐𝑜𝑢𝑛𝑡

𝐶𝑜𝑟𝑟𝑒𝑐𝑡𝑒𝑑 𝑊𝐵𝐶 𝑐𝑜𝑢𝑛𝑡 = 𝑥 100
100 + # 𝑜𝑓 𝑁𝑅𝐵𝐶𝑠/100 𝑊𝐵𝐶𝑠

Reference:
Keohane, E., Otto, C., & Walenga, J. (2016). Rodak’s Hematology Clinical Principles and Applications (6th
ed). Elsevier Inc. ISBN: 978-0-323-53045-3

4. Describe proper way of cleaning and drying (Hematology) diluting pipettes (4 points)

1) Attach the end of the pipette with the suction (pump) device.
2) Fill the pipette with water and thoroughly wash out the diluted blood until traces are
removed.
3) Draw and expel a bleach solution for a few times, followed by a rinse of distilled
water to clean the pipette from stains.
4) Rinse the pipette with absolute (95%) alcohol to remove the water within, then, use
either acetone or ether to remove alcohol.
5) Pass air through to dry the pipette, so that the bead rolls freely (free movement) in
the bulb.

** It is a very laborious process to clean the pipettes by blowing through them, further, one
cannot dry them properly by blowing, as expired air is laden with moisture.

References:
AUN Edu. (n.d.). III. Errors in counting total RBCs and WBCs by haemocytometer.
www.aun.edu.eg/developmentvet/clinical%20laboratory%20diagnosis/D_1_c.htm.
Klimud. (n.d.). Fundamentals of Hematology Cell Counts and Measurements. https://www.klim
ud.org/public/atlas/idrar/web/www.irvingcrowley.com/cls/fund.htm?fbclid=IwAR325UZ
nkfgUgghg2cXnYVkIMtNOtSEkSvnJ5gxHmsbST0xpin5UmQ3mpeo
5. Give 5 sources of errors in manual hemocytometer (cell counting). Briefly explain each (10
points)

1) Not Differentiating Between Cells and Debris

Even to the highly-trained eye, sometimes debris can come across as just another cell in
the sample. There is a chance that debris will be present, and when this debris is
counted as a cell it is known as a false-positive. Sometimes, manual cell counters will
also misclassify a cell as debris, resulting in a false-negative.

2) Volume, Dilution, and Pipetting Errors

Though the hemocytometer contains a given volume, the space between the counting
chamber and the cover glass might be slightly increased when the chamber is filled with
liquid. This can result in an underestimation of the sample volume causing
overestimation of cell concentration, leading to errors based on estimating the volume
incorrectly.

3) Not Suspending a Sample Properly

When a cell suspension rests, many of the cells in the suspension will move toward the
bottom of the test tube. A sample taken from this tube will not represent the true
solution and result in inaccurate cell values.

4) Human Subjectivity
Each person performing the manual cell count adheres to a certain set of criteria that
defines a cell along with the stain intensity threshold to count it as viable or dead. Such
variations in human perception when counting manually can be extremely detrimental
to experimental setup and analysis when counting cells manually. If multiple users
count the same sample, it is not uncommon to see a variance significantly higher than
the mean of a Poisson distribution.

5) Mathematical Errors
With any type of data collection, a good deal of math is involved. The use of
mathematics is essential in calculating the average number of cells in a suspension.
Math performed manually is time consuming and is also error-prone, as it relies on the
skill of the person involved. Improper formulas, miscomputations, and typographical
errors like mistyped numbers and mathematical functions, can lead to huge
inaccuracies in the values of the cells.

References:
Chemometec. (n.d.). Manual vs. Automated Cell Counting. https://chemometec.com/resourc
es/mini-reviews/manual-vs-automated-cell-counting/
Corning. (n.d.). 10 Most Common Errors Made in Cell Counting. https://www.corning.com/cata
log/cls/documents/application-notes/CLS-AN-495.pdf