Topic Introduction

Fluorescence Microscopy

  1. Martin D. Bootman3
  1. 1Department of Microbiology and Physiology Systems, University of Massachusetts Medical School, Worcester, Massachusetts 01655;
  2. 2Department of Neurobiology & Behavior, University of California, Irvine, California 92697-4550;
  3. 3Department of Life, Health and Chemical Sciences, The Open University, Milton Keynes MK7 6AA, United Kingdom

    Abstract

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation.

    Footnotes

    • ↵4 Correspondence: michael.sanderson{at}umassmed.edu

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    1. doi:10.1101/pdb.top071795 Cold Spring Harb Protoc 2014: pdb.top071795- © 2014 Cold Spring Harbor Laboratory Press
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