[Preparation of chicken red blood cells for calibration of flow cytometry].
OBJECTIVE
To prepare stable chicken red blood cells for the calibration of flow cytometry.


METHODS
The traditional isolation method of chicken red blood cells was modified by incorporating gelatin technique, Ca2+-free HBSS treatment and low-speed centrifugation. The effect of fluorescence staining of the cells was improved by the addition of TritonX-100 to enhance the membrane permeability and Rnase enzymes to disintegrate RNA tiles. The modified method was compared with the traditional method for viability of the freshly isolated cells and the DNA content coefficient of variation (CV) of the fixed cells.


RESULTS
Chicken red blood cells obtained by the modified method showed a significantly higher viability than those obtained by the traditional method [(98.5∓3.5)% vs (93.5∓2.7)%, P<0.05]. After glutaraldehyde fixation, the isolated cells with the modified method were stable during the 90-day preservation with a significantly lower CV than the cells obtained by the traditional method [(6.0∓0.3)% to 6.2∓0.4% vs (8.6∓0.5)% to (13.1∓1.4)%, P<0.01].


CONCLUSIONS
The chicken red blood cells isolated using the modified method can be applicable for calibration of flow cytometry.
Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China. yinj@sibet.ac.cn
Jian
J
Yin
Shutao
S
Zhao
Xiaodong
X
Wu
Ce
C
Wang
Yunliang
Y
Wu
D000818:Animals; D002138:Calibration; D002645:Chickens; D004912:Erythrocytes / Q000166:cytology*; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*