Actually, I am working on colorimetric analysis of urine analyte. In my analysis, I need to prepare samples of 10, 20, 50 and 100 RBC per micoliter from from collected blood to react with urine test strip and study its color change for further analysis. We do not have hematocytometer or any hematology analyzer. Is there any straight forward method to prepare those RBC samples from the collected blood?
Dear Jalal,
check if these links, paper or references are of any help to you:
https://labtestsonline.org/understanding/analytes/rbc/tab/sample/
https://www.thermofisher.com/fi/en/home/references/protocols/cell-and-tissue-analysis/elisa-protocol/elisa-sample-preparation-protocols/plasma-and-serum-preparation.html#
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Dear Jalal
in the strips for urine analysis not RBC number but the presence of hemoglobin at different concentrations are measured. This because in the specific "blood area" of the strip, the lysis of RBC that are present in the sample occurs. I think it's very difficult to obtain a "calibration curve" with 10, 20, 50, 100 RBC without any counting system! Anyway, considering that a "normal blood sample" could contains 4.5 - 5.5 milions of RBC, in 20microL there are 90-110 milions of RBC. By diluting 20microL of blood in 1L of saline you obtain a blood solution with the final concentration ranging from 90 to 110 RBC/microL.
Collect your blood with EDTA or CPD , wash 3 times at 1500 rpm and dilute your blood sample at 1ml of PBS. Then , count the number of cells by hemocytometer and separate the number of cells accordingly your plan.
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