Flow Cytometric Method for Counting Residual Leukocytes.
- Author:
Kyeong Lan CHOI
1
;
Duck CHO
;
Jong Hee SHIN
;
Soon Pal SUH
;
Dong Wook RYANG
Author Information
1. Gwangju-Jeonnam Red Cross Blood Center, Korea. choiorchid@yahoo.com
- Publication Type:Original Article
- Keywords:
Residual leukocytes;
Flow cytometer;
PI;
Blood components
- MeSH:
DNA;
Leukocytes*;
Limit of Detection;
Propidium
- From:Korean Journal of Blood Transfusion
2004;15(1):1-7
- CountryRepublic of Korea
- Language:Korean
-
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.
