Ohn, Jungho and Yang, Jennifer and Fraser, Scott E. and Lansford, Rusty and Liebling, Michael (2011) High-speed multicolor microscopy of repeating dynamic processes. Genesis, 49 (7). pp. 514-521. ISSN 1526-954X http://resolver.caltech.edu/CaltechAUTHORS:20110801-093521155
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Images of multiply labeled fluorescent samples provide unique insights into the localization of molecules, cells, and tissues. The ability to image multiple channels simultaneously at high speed without cross talk is limited to a few colors and requires dedicated multichannel or multispectral detection procedures. Simpler microscopes, in which each color is imaged sequentially, produce a much lower frame rate. Here, we describe a technique to image, at high frame rate, multiply labeled samples that have a repeating motion. We capture images in a single channel at a time over one full occurrence of the motion then repeat acquisition for other channels over subsequent occurrences. We finally build a high-speed multichannel image sequence by combining the images after applying a normalized mutual information-based time registration procedure. We show that this technique is amenable to image the beating heart of a double-labeled embryonic quail in three channels (brightfield, yellow, and mCherry fluorescent proteins) using a fluorescence wide-field microscope equipped with a single monochrome camera and without fast channel switching optics. We experimentally evaluate the accuracy of our method on image series from a two-channel confocal microscope.
|Additional Information:||© 2011 Wiley-Liss, Inc. Received 18 December 2010; Revised 18 April 2011; Accepted 30 May 2011. Article first published online: 21 Jun. 2011. Contract grant sponsor: National Institutes of Health, Contract grant number: HL095586 (to M.L.).|
|Subject Keywords:||cardiac imaging; quail; fluorescence imaging; multicolor imaging; heart development; registration; fast imaging; mutual information|
|Official Citation:||Ohn, J., Yang, J., Fraser, S. E., Lansford, R. and Liebling, M. (2011), High-speed multicolor microscopy of repeating dynamic processes. genesis, 49: 514–521. doi: 10.1002/dvg.20774|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Tony Diaz|
|Deposited On:||14 Oct 2011 20:34|
|Last Modified:||14 Oct 2011 20:34|
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