Open AccessComparison of the automated fluorescence microscopic viability test with the conventional and flow cytometry methods
Author(s) -
Kim Jang Su,
Nam Myung Hyun,
An Seong Soo A.,
Lim Chae Seung,
Hur Dae Sung,
Chung Chanil,
Chang Jun Koon
Publication year - 2011
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.20438
Subject(s) - trypan blue , flow cytometry , propidium iodide , cytometry , viability assay , biomedical engineering , cell counting , stain , staining , chemistry , chromatography , materials science , cell , microbiology and biotechnology , pathology , medicine , biology , cell cycle , biochemistry , apoptosis , programmed cell death
The cell viability test is an essential tool in any laboratory, performing cell‐based studies and clinical laboratory tests. The trypan blue exclusion method is the most popular assay for its simple concept among various diagnostic tools. However, several disadvantages include time‐consuming and labor‐intensive steps with low precision. In this study, we evaluated a new technique for the automatic cell viability measurement with microscopic cell counter and microchip. Upon blood draw from 11 healthy volunteers, Mononuclear cells were separated immediately from the heparinized whole blood, and the viable cells were diluted from 100 to 1%. The cell viability tests were performed simultaneously with following three methods: the conventional manual trypan blue exclusion method; the flow cytometry measurement with propidium iodide stain; and the newly developed microscopic cell counter with microchip. Linearities, precisions, and correlations from three methods were analyzed and compared. The correlations data from the microscopic cell counter were in good agreement with both the conventional trypan blue method ( r =0.99, P <0.05) and the flow cytometry ( r =0.99, P <0.05), respectively. The precision (2.0–6.2%) and linearity from the microscopic cell counter method with microchip were superior in comparison with the conventional method. The microscopic cell counter with microchip performed well with high precision, linearity, and efficient running time than both the manual trypan blue and the flow cytometry methods. J. Clin. Lab. Anal. 25:90–94, 2011. © 2011 Wiley‐Liss, Inc.