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Methods Cell Biol. 2014;123:193-215. doi: 10.1016/B978-0-12-420138-5.00011-2.

Light sheet microscopy.

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1
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.

Abstract

This chapter introduces the concept of light sheet microscopy along with practical advice on how to design and build such an instrument. Selective plane illumination microscopy is presented as an alternative to confocal microscopy due to several superior features such as high-speed full-frame acquisition, minimal phototoxicity, and multiview sample rotation. Based on our experience over the last 10 years, we summarize the key concepts in light sheet microscopy, typical implementations, and successful applications. In particular, sample mounting for long time-lapse imaging and the resulting challenges in data processing are discussed in detail.

KEYWORDS:

Developmental biology; Fluorescence microscopy; Image processing; Light sheet microscopy; Mounting; Selective plane illumination microscopy; Three-dimensional imaging; Ultramicroscopy; Zebrafish

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