Fluorescent probes and flow cytometry to assess rat sperm integrity and mitochondrial function

Fluorescent assessment of cellular integrity and mitochondrial function by flow cytometry can provide a rapid and precise means of determining the fun ctional status of large numbers of spermatozoa. In the present study, rat s perm viability was assessed with SYBR-14 and PI and sperm mitochondria were differentially labeled with JC-1. Sperm samples of variable viability were prepared using varying proportions of fresh and frozen spermatozoa. SYBR-1 4 stained sperm correlated well with expected sperm viability (r = 0.98). M otile sperm stained with JC-1 appeared orange in the midpiece indicating a high mitochondrial membrane potential whereas: immotile sperm with a low me mbrane potential stained green. The percentage of spermatozoa staining oran ge was highly correlated (r = 0.99) with expected sperm viability. Flow cyt ometry using specific fluorescent probes is a useful technique fur detectin g changes in rat sperm plasma membrane integrity and mitochondrial function in large numbers of spermatozoa. (C) 2001 Elsevier Science Inc. All rights reserved.