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Bio Protoc. 2019 Mar 5;9(5). pii: e3182. doi: 10.21769/BioProtoc.3182.

Single-cell Motility Analysis of Tethered Human Spermatozoa.

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
2
Department of Urology, University of California, San Francisco, CA, USA.
3
Gladstone Institute of Virology and Immunology, San Francisco, CA, USA.

Abstract

Vigorous sperm flagellar motility is essential for fertilization, and so the quantitative measurement of motility is a useful tool to assess the intrinsic fertility potential of sperm cells and explore how various factors can alter sperm's ability to reach the egg and penetrate its protective layers. Human sperm beat their flagella many times each second, and so recording and accurately quantifying this movement requires a high-speed camera. The aim of this protocol is to provide a detailed description of the tools required for quantitative beat frequency measurement of tethered human sperm at the single-cell level and to describe methods for investigating the effects of intracellular or extracellular factors on flagellar motion. This assay complements bulk measurements of sperm parameters using commercially-available systems for computer-assisted sperm analysis (CASA).

KEYWORDS:

Amyloid fibrils; Flagellar beat frequency; High-speed imaging; Human spermatozoa; Seminal plasma; Single-cell motility analysis; Sperm Swim-up; Sperm flagellum; Sperm motility

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