High-speed multicolor microscopy of repeating dynamic processes.

High-speed multicolor microscopy of repeating dynamic processes.
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重复动态过程的高速多色显微镜。

DOI:
10.1002/dvg.20774
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发表时间:
2011
期刊:
Genesis (New York, N.Y. : 2000)
影响因子:
--
通讯作者:
Liebling,Michael
Liebling,Michael
中科院分区:
--
文献类型:
--
作者:
Ohn,Jungho;Yang,Jennifer;Fraser,ScottE;Lansford,Rusty;Liebling,Michael

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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. genesis 49:514–521, 2011. © 2011 Wiley‐Liss, Inc.
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