High throughput, parallel imaging and biomarker quantification of human spermatozoa by ImageStream flow cytometry.
Spermatid specific thioredoxin-3 protein (SPTRX-3) accumulates in the superfluous cytoplasm of defective human spermatozoa. Novel ImageStream technology combining flow cytometry with cell imaging was used for parallel quantification and visualization of SPTRX-3 protein in defective spermatozoa of five men from infertile couples. The majority of the SPTRX-3 containing cells were overwhelmingly spermatozoa with a variety of morphological defects, detectable in the ImageStream recorded images. Quantitative parameters of relative SPTRX-3 induced fluorescence measured by ImageStream correlated closely with conventional flow cytometric measurements of the same sample set and reflected the results of clinical semen evaluation. Image Stream quantification of SPTRX-3 combines and surpasses the informative value of both conventional flow cytometry and light microscopic semen evaluation. The observed patterns of the retention of SPTRX-3 in the sperm samples from infertility patients support the view that SPTRX3 is a biomarker of male infertility.
Buckman
Clayton
C
Division of Animal Sciences, University of Missouri, Columbia, MO 65211-5300, USA.
George
Thaddeus C
TC
Friend
Sherree
S
Sutovsky
Miriam
M
Miranda-Vizuete
Antonio
A
Ozanon
Christophe
C
Morrissey
Phil
P
Sutovsky
Peter
P
D000328:Adult; D015415:Biomarkers / Q000032:analysis*; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis; D008297:Male; D013094:Spermatozoa / Q000648:ultrastructure*; D013879:Thioredoxins / Q000032:analysis*
D015415:Biomarkers; C490377:TXNDC8 protein, human; D013879:Thioredoxins