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Proceedings Paper

A real-time single sperm tracking, laser trapping, and ratiometric fluorescent imaging system
Author(s): Linda Z. Shi; Elliot L. Botvinick; Jaclyn Nascimento; Charlie Chandsawangbhuwana; Michael W. Berns
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Paper Abstract

Sperm cells from a domestic dog were treated with oxacarbocyanine DiOC2(3), a ratiometrically-encoded membrane potential fluorescent probe in order to monitor the mitochondria stored in an individual sperm's midpiece. This dye normally emits a red fluorescence near 610 nm as well as a green fluorescence near 515 nm. The ratio of red to green fluorescence provides a substantially accurate and precise measurement of sperm midpiece membrane potential. A two-level computer system has been developed to quantify the motility and energetics of sperm using video rate tracking, automated laser trapping (done by the upper-level system) and fluorescent imaging (done by the lower-level system). The communication between these two systems is achieved by a networked gigabit TCP/IP cat5e crossover connection. This allows for the curvilinear velocity (VCL) and ratio of the red to green fluorescent images of individual sperm to be written to the hard drive at video rates. This two-level automatic system has increased experimental throughput over our previous single-level system (Mei et al., 2005) by an order of magnitude.

Paper Details

Date Published: 30 August 2006
PDF: 8 pages
Proc. SPIE 6326, Optical Trapping and Optical Micromanipulation III, 63260V (30 August 2006); doi: 10.1117/12.680912
Show Author Affiliations
Linda Z. Shi, Univ. of California, San Diego (United States)
Elliot L. Botvinick, Univ. of California, Irvine (United States)
Jaclyn Nascimento, Univ. of California, San Diego (United States)
Charlie Chandsawangbhuwana, Univ. of California, San Diego (United States)
Michael W. Berns, Univ. of California, San Diego (United States)
Univ. of California, Irvine (United States)


Published in SPIE Proceedings Vol. 6326:
Optical Trapping and Optical Micromanipulation III
Kishan Dholakia; Gabriel C. Spalding, Editor(s)

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