Journal of Medical Diagnostic Methods
Open Access
ISSN: 2168-9784
+44 1300 500008
ISSN: 2168-9784
+44 1300 500008
Laboratory of Pathology and Development, Institute for Quantitative Biosciences, The University of Tokyo, 1-1 Yayoi, Bunkyo, Tokyo, 113-0032, Japan
Research Article
Sperm Specific Two-Step Dye Exclusion Assays to Evaluate Integrity of Plasma and Organelle Membranes-New Approach for Quality Assurance of the Sperm for Intra-Cytoplasmic Sperm Injection
Author(s): Satoru Kaneko*, Yuki Okada and Kiyoshi Takamatsu
Objective: The comprehensive assessments of the plasma and organelle membranes are new approach to understand human sperm fertility. The present study developed 2 types of the sperm specific 2-step dye exclusion assays to evaluate the plasma and organelle membranes in human sperm.
Methods: Sperm specific 2-step dye exclusion assay: Sperm was incubated with reactive red 195 (RR195) or Cy3- conjugated concanavalin A (Cy3-con A) in isotonic culture medium, they were counterstained with reactive blue 222 or Alexa 488 conjugated concanavalin A after exclusion of the plasma membrane with methanol, respectively. Integrity of the organelle membrane in the mitochondria was observed by retention of Mito Tracker FM. The cellROX Orange produced orange fluorescence upon oxidation by reacted reactive oxygen species.
Results: The.. View More»
DOI:
10.35248/2168-9784.23.12.450