Light-Sheet Fluorescence Microscopy

Yang Yulong1 Zong Weijian1 Wu Runlong1 Chen Liangyi1

(1.State Key Laboratory of Membrane Biology, Institute of Molecular Medicine, Peking University, Beijing, China 100871)

【Abstract】In the past two decades, laser scanning confocal microscope has been the standard tool for observing the process of life at cellular and sub-cellular level. The optical sectioning capacity of pinhole-based confocal microscope comes at the price of unwanted excitation of fluorophores out of focal plane and phototoxic damage to biological samples. As a new type of fluorescent microscope, light-sheet fluorescence microscope (LSFM) uses side illumination to conduct surface imaging of the samples directly. As compared to the point-scanning imaging mode, LSFM excels at its imaging speed, which is much higher than that of laser scanning confocal microscope, thus making it possible to study some high-speed fine life activities. Another advantage of the light-sheet fluorescence microscope is that only the sample at the light sheet is excited and the sample outside the light sheet is not excited, so there is less phototoxic dosage and we can observe the sample in a longer time scale. The special illumination and imaging mode of the light-sheet fluorescence microscope make it play an irreplaceable role in three-dimensional high-speed imaging of big biological samples. The history and research status of light-sheet fluorescence microscope are reviewed with the purpose of providing a personal perspective of current situation and future direction of LSFM.

【Keywords】 microscopy; fluorescence imaging; light-sheet illumination; micro-imaging; phototoxicity;

【DOI】

【Funds】 Fund for National Key Research and Development Project (2016YFA0500400) National Basic Research Program of China (973 Program) (2013CB531200) National Natural Science Foundation of China (31327901, 31521062, 31570839)

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(Translated by caizhijian)

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This Article

ISSN:0253-2239

CN: 31-1252/O4

Vol 37, No. 03, Pages 71-78

March 2017

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Article Outline

Abstract

  • 1 Introduction
  • 2 Selective plane illumination microscope with layered light
  • 3 Digital scanning light-sheet microscope
  • 4 High-resolution light-sheet microscope
  • 5 Application and future prospects of light-sheet microscope
  • References