[Evaluation of sperm plasma membrane integrity by SYBR-14/PI dual fluorescent staining and flow cytometry].
OBJECTIVE
To investigate the feasibility and clinical significance of detecting the plasma membrane integrity (PMI) of sperm by SYBR-14/PI fluorescent staining and flow cytometry.


METHODS
A total of 208 semen samples were divided into a normal (n = 31) and an abnormal group (n = 177), subjected to conventional computer-assisted semen analysis (CASA), and then evaluated for sperm PMI by flow cytometry after washed and SYBR-14/PI dual fluorescent staining. The percentage of sperm with PMI was indicated as that of the sperm emitting green fluorescence (SYBR-14+/PI- %).


RESULTS
Significant differences were detected in SYBR-14+/PI- and SYBR-14-/PI+ between the normal and abnormal groups (P < 0.05), with the SYBR-14+/PI- % significantly higher in the former ([55.66 +/- 20.64] %) than in the latter ([39.71 +/- 19.21] %) (P = 0.000). The SYBR-14+/PI-% showed significant positive correlations with sperm motility (r = 0.408, P = 0.000) and the percentage of grade a + b sperm (r = 0.398, P = 0.000), and a significant negative correlation with the percentage of grade d sperm (r = -0.413, P = 0.000); the SYBR-14-/PI+ % exhibited significant negative correlations with sperm motility (r = -0.380, P = 0.000) and the percentage of grade a + b sperm (r = -0.397, P = 0.000), and a significant positive correlation with the percentage of grade d sperm (r = 0.385, P = 0.000); the SYBR-14+/PI+ % showed positive correlations with sperm motility (r = 0.172, P = 0.013) and the percentage of grade a + b sperm (r = 0.177, P = 0.011), and a negative correlation with grade d sperm (r = -0.164, P = 0.018).


CONCLUSIONS
SYBR-14/PI dual fluorescent staining and flow cytometry could be readily used to detect sperm PMI and evaluate male reproductivity.
Zhou
Xin
X
College of Life Science, Nanjing Normal University, Nanjing, Jiangsu 210046, China.
Xia
Xin-Yi
XY
Cao
Qi
Q
Kang
Ning
N
Fan
Xiao-Bo
XB
Shao
Yong
Y
Li
Ying
Y
Huang
Yu-Feng
YF
D000328:Adult; D016022:Case-Control Studies; D002462:Cell Membrane; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D009930:Organic Chemicals; D013081:Sperm Motility; D013094:Spermatozoa
D005456:Fluorescent Dyes; D009930:Organic Chemicals; C093207:SYBR-14