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      Single-molecule fluorescence microscopy review: shedding new light on old problems

      1 , 2 , , 1 , 2

      Bioscience Reports

      Portland Press Ltd.

      fluorescence, microscopy, single-molecule, super-resolution

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          Abstract

          Fluorescence microscopy is an invaluable tool in the biosciences, a genuine workhorse technique offering exceptional contrast in conjunction with high specificity of labelling with relatively minimal perturbation to biological samples compared with many competing biophysical techniques. Improvements in detector and dye technologies coupled to advances in image analysis methods have fuelled recent development towards single-molecule fluorescence microscopy, which can utilize light microscopy tools to enable the faithful detection and analysis of single fluorescent molecules used as reporter tags in biological samples. For example, the discovery of GFP, initiating the so-called ‘green revolution’, has pushed experimental tools in the biosciences to a completely new level of functional imaging of living samples, culminating in single fluorescent protein molecule detection. Today, fluorescence microscopy is an indispensable tool in single-molecule investigations, providing a high signal-to-noise ratio for visualization while still retaining the key features in the physiological context of native biological systems. In this review, we discuss some of the recent discoveries in the life sciences which have been enabled using single-molecule fluorescence microscopy, paying particular attention to the so-called ‘super-resolution’ fluorescence microscopy techniques in live cells, which are at the cutting-edge of these methods. In particular, how these tools can reveal new insights into long-standing puzzles in biology: old problems, which have been impossible to tackle using other more traditional tools until the emergence of new single-molecule fluorescence microscopy techniques.

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          Author and article information

          Journal
          Biosci Rep
          Biosci. Rep
          ppbioscirep
          BSR
          Bioscience Reports
          Portland Press Ltd.
          0144-8463
          1573-4935
          10 July 2017
          21 July 2017
          31 August 2017
          : 37
          : 4
          Affiliations
          [1 ]Department of Physics, Biological Physical Sciences Institute (BPSI), University of York, York YO10 5DD, U.K.
          [2 ]Department of Biology, Biological Physical Sciences Institute (BPSI), University of York, York YO10 5DD, U.K.
          Author notes
          Correspondence: Mark C. Leake ( mark.leake@ 123456york.ac.uk )
          Article
          10.1042/BSR20170031
          5520217
          28694303
          © 2017 The Author(s).

          This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY).

          Page count
          Pages: 19
          Product
          Categories
          Review Articles
          Review Article

          Life sciences

          fluorescence, super-resolution, single-molecule, microscopy

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