title: Wide-field four-channel fluorescence imager for biological applications.
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 x 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.
authors.affiliation: Rice University, Department of Chemical and Biomolecular Engineering, 6100 South Main Street, Houston, Texas 77005, USA.
authors.forename: Madhuri
authors.initials: M
authors.lastname: Thakur
authors.forename: Dmitry
authors.initials: D
authors.lastname: Melnik
authors.forename: Heather
authors.initials: H
authors.lastname: Barnett
authors.forename: Kevin
authors.initials: K
authors.lastname: Daly
authors.forename: Christine H
authors.initials: CH
authors.lastname: Moran
authors.forename: Wei-Shun
authors.initials: WS
authors.lastname: Chang
authors.forename: Stephan
authors.initials: S
authors.lastname: Link
authors.forename: Christopher Theodore
authors.initials: CT
authors.lastname: Bucher
authors.forename: Carter
authors.initials: C
authors.lastname: Kittrell
authors.forename: Robert
authors.initials: R
authors.lastname: Curl
mesh_terms: D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008029:Lighting / Q000295:instrumentation*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity