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Journal of Biomedical Optics

Use of laser tweezers to analyze sperm motility and mitochondrial membrane potential
Author(s): Jaclyn M. Nascimento; Linda Z. Shi; Charlie Chandsawangbhuwana; James Tam; Barbara S. Durrant; Elliot L. Botvinick; Michael W. Berns
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Paper Abstract

We combine laser tweezers with custom computer tracking software and robotics to analyze the motility [swimming speed, VCL (curvilinear velocity), and swimming force in terms of escape laser power (Pesc)] and energetics [mitochondrial membrane potential (MP)] of individual sperm. Domestic dog sperm are labeled with a cationic fluorescent probe, DiOC2(3), that reports the MP across the inner membrane of the mitochondria located in the sperm's midpiece. Individual sperm are tracked to calculate VCL. Pesc is measured by reducing the laser power after the sperm is trapped using laser tweezers until the sperm is capable of escaping the trap. The MP is measured every second over a 5-s interval during the tracking phase (sperm is swimming freely) and continuously during the trapping phase. The effect of the fluorescent probe on sperm motility is addressed. The sensitivity of the probe is measured by assessing the effects of a mitochondrial uncoupling agent (CCCP) on MP of free swimming sperm. The effects of prolonged exposed to the laser tweezers on VCL and MP are analyzed. The system's capabilities are demonstrated by measuring VCL, Pesc, and MP simultaneously for individual sperm. This combination of imaging tools is useful to quantitatively assess sperm quality and viability.

Paper Details

Date Published: 1 January 2008
PDF: 7 pages
J. Biomed. Opt. 13(1) 014002 doi: 10.1117/1.2839051
Published in: Journal of Biomedical Optics Volume 13, Issue 1
Show Author Affiliations
Jaclyn M. Nascimento, Univ. of California/San Diego (United States)
Linda Z. Shi, Univ. of California/San Diego (United States)
Charlie Chandsawangbhuwana, Univ. of California/San Diego (United States)
James Tam, Univ. of California/San Diego (United States)
Barbara S. Durrant, Conservation and Research for Endangered Species (United States)
Elliot L. Botvinick, Univ. of California/Irvine (United States)
Michael W. Berns, Univ. of California/Irvine (United States)

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