A real-time single sperm tracking, laser trapping, and ratiometric fluorescent imaging system

Linda Z. Shi; Elliot L. Botvinick; Jaclyn Nascimento; Charlie Chandsawangbhuwana; Michael W. Berns

doi:10.1117/12.680912

30 August 2006 A real-time single sperm tracking, laser trapping, and ratiometric fluorescent imaging system

Linda Z. Shi, Elliot L. Botvinick, Jaclyn Nascimento, Charlie Chandsawangbhuwana, Michael W. Berns

Proceedings Volume 6326, Optical Trapping and Optical Micromanipulation III; 63260V (2006) https://doi.org/10.1117/12.680912
Event: SPIE Optics + Photonics, 2006, San Diego, California, United States

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.

Citation Download Citation

Linda Z. Shi, Elliot L. Botvinick, Jaclyn Nascimento, Charlie Chandsawangbhuwana, and Michael W. Berns "A real-time single sperm tracking, laser trapping, and ratiometric fluorescent imaging system", Proc. SPIE 6326, Optical Trapping and Optical Micromanipulation III, 63260V (30 August 2006); https://doi.org/10.1117/12.680912

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available