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Journal of Biomedical Optics • Open Access

Wide-field four-channel fluorescence imager for biological applications
Author(s): Madhuri M. Thakur; Dmitry G. Melnik; Heather Barnett; Kevin Daly; Christine H. Moran; Wei-Shun Chang; Stephan Link; C. Theodore Bucher; Carter Kittrell; Robert F. Curl

Paper Abstract

A wide-field four-channel fluorescence imager has been developed. The instrument uses four expanded laser beams to image a large section (6 mm×9 mm). An object can be sequentially illuminated with any combination of 408-, 532-, 658-, and 784-nm lasers for arbitrary (down to 1 ms) exposure times for each laser. Just two notch filters block scattered light from all four lasers. The design approach described here offers great flexibility in treatment of objects, very good sensitivity, and a wide field of view at low cost. There appears to be no commercial instrument capable of simultaneous fluorescence imaging of a wide field of view with four-laser excitation. Some possible applications are following events such as flow and mixing in microchannel systems, the transmission of biological signals across a culture, and following simulations of biological membrane diffusion. It can also be used in DNA sequencing by synthesis to follow the progress of the photolytic removal of dye and terminator. Without utilizing its time resolution, it can be used to obtain four independent images of a single tissue section stained with four targeting agents, with each coupled to a different dye matching one of the lasers.

Paper Details

Date Published: 1 March 2010
PDF: 9 pages
J. Biomed. Opt. 15(2) 026016 doi: 10.1117/1.3374052
Published in: Journal of Biomedical Optics Volume 15, Issue 2
Show Author Affiliations
Madhuri M. Thakur, Rice Univ. (United States)
Dmitry G. Melnik, The Ohio State Univ. (United States)
Heather Barnett, Univ. of Washington (United States)
Kevin Daly, Princeton Univ. (United States)
Christine H. Moran, Washington Univ. in St. Louis (United States)
Wei-Shun Chang, Rice Univ. (United States)
Stephan Link, Rice Univ. (United States)
C. Theodore Bucher, Rice Univ. (United States)
Carter Kittrell, Rice Univ. (United States)
Robert F. Curl, Rice Univ. (United States)

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