Korean J Blood Transfus.  2004 Jun;15(1):1-7.

Flow Cytometric Method for Counting Residual Leukocytes

• Choi KL
• Cho D
• Shin JH
• Suh SP
• Ryang DW

Abstract

BACKGROUND: Simple manual method using a Nageotte hemocytometer counting residual leukocytes [White blood cells(WBC)] in blood components is subjective, labor-intensive, time consuming and variable within and between laboratories. The aim of this study was to evaluate usefulness for flow cytometric method to determinate very low numbers of leukocytes in leukocyte-free blood components.
METHODS
Epics XL-MCL (Beckman Coulter Co.) was used for determination of fluorescence-labelled cells. The DNA in leukocytes was stained using propidium iodide (PI) and leukocytes were automatically analysed by flow cytometer.
RESULTS
Linearity determined over a range of 0.5-50 WBC/muL was a value of r=0.994. The detection limit of this method was 4.4 WBC/muL and accuracy was 86.6% with linearity of r=0.991 over the 5-50 WBC/muL. Reproducibility was CV of 9.1% for 25.8 WBC/muL and 14.7% for 7.1 WBC/muL, respectively.
CONCLUSION
Flow cytometric techniques provide a reproducible and objective tool for counting residual WBC in leukocyte free blood components compared with the Nageotte hemocytometer.

Keyword

Residual leukocytes; Flow cytometer; PI; Blood components

MeSH Terms

DNA
Leukocytes*
Limit of Detection
Propidium
DNA
Propidium