A novel in vitro sperm head decondensation protocol for rapid flow cytometric measurement of deoxyribonucleic acid content  

Oleh:

Antonucci, Niki ; Manes, Sabrina ; Corradetti, Bruna ; Manicardi, Gian Carlo ; Borini, Andrea ; Bizzaro, Davide

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Fertility and Sterility (keterangan: ada di ClinicalKey) vol. 99 no. 07 (Jun. 2013), page 1857-1861.
Topik: Sperm decondensation protocol; flow cytometry; DNA content; sperm chromatin packaging

Ketersediaan

Perpustakaan FK Nomor Panggil: F02.K.2013.04 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

Objective To set up a novel protocol of sperm head in vitro decondensation that obviates the problematic effect of the variable degree of sperm chromatin packaging on DNA staining needed for flow cytometric analysis. Design Development of a new cytofluorimetric assay. Setting University laboratory. Patient(s) Semen specimens were obtained from normospermic healthy volunteers at the Department of Life and Environmental Sciences, Università Politecnica delle Marche. Intervention(s) Setup of the novel in vitro sperm head decondensation protocol; sperm were then stained and analyzed by flow cytometry to measure DNA content. Main Outcome Measure(s) Mean fluorescent channel, DNA content, percentage diploid sperm. Result(s) Native nondecondensed fluorochrome-labeled sperm show significant under-staining, resulting in an underestimated C-value (approximately 1.4 pg). This protocol ensures stoichiometric staining of sperm DNA, which becomes fully reachable by fluorescent probes and makes the diploid (7.12 pg) over haploid (3.56 pg) sperm frequency quantification easier. Conclusion(s) This study establishes a simple method for in vitro sperm head decondensation, which allows accurate detection of the real sperm DNA content.

Opini Anda

Klik untuk menuliskan opini Anda tentang koleksi ini!

Kembali