Single-molecule imaging’s insights into the puzzling components of cells

Iva Tolić-Nørrelykke and colleagues at the Max Planck Institute of Molecular Cell Biology and Genetics, Germany, have written a fascinating tutorial review on how single-molecule imaging is revealing the pieces of cellular jigsaw puzzles.

single-molecule imaging

They see the cell as a dynamic puzzle, with pieces being created, destroyed and changing their interactions all the time. These pieces can now be viewed in vivo using various techniques in single-molecule microscopy with high precision. The measurement of intracellular reactions can also be carried out in order to build models of intricate cellular processes.

As well as a brief chronological look at the developments in single-molecule microscopy and useful short explanations of the different techniques as a glossary, this review includes: 

  1. Appropriate labelling methods
  2. Total Internal Reflection Fluorescence (TIRF) in vivo
  3. Widefield microscopy for a low signal-to-noise ratio and Selective Plane Illumination Microscopy (SPIM)
  4. Fluorescence Speckle Microscopy using tubulin and Photoactivation with a specific wavelength
  5. Mapping lots of individual molecules inside a cell using super-resolution fluorescence methods
  6. Single-molecule FRET for studying conformational change

They emphasise that due to the complications arising inside a cell, the molecules must be resolved spatially in different ways according to the different techniques and they discuss which techniques are best suited to which types of investigation. Single-molecule imaging is compared with ensemble methods and the goal of measuring a large number of molecular events simultaneously in real time is discussed.

Learn more about the potential of single-molecule imaging in the full Tutorial review:

Single-molecule imaging in vivo: the dancing building blocks of the cell
Miguel Coelho, Nicola Maghelli and Iva M. Tolić-Nørrelykke
DOI: 10.1039/C3IB40018B

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