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.
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:
- Appropriate labelling methods
- Total Internal Reflection Fluorescence (TIRF) in vivo
- Widefield microscopy for a low signal-to-noise ratio and Selective Plane Illumination Microscopy (SPIM)
- Fluorescence Speckle Microscopy using tubulin and Photoactivation with a specific wavelength
- Mapping lots of individual molecules inside a cell using super-resolution fluorescence methods
- 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