Probing the future of correlative microscopy
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EDITORIAL
Probing the future of correlative microscopy Lucy Collinson 1 & Paul Verkade 2
Published online: 30 September 2015 # Springer-Verlag Berlin Heidelberg 2015
Instrumental developments in microscopy, such as confocal scanning light microscopy and super resolution light microscopy, have firmly established imaging as a key technology in modern life science research. However, this would not have been possible without the availability of the right probes—the concurrent emergence of green fluorescent protein (GFP) completely transformed the field. We can surely assume that GFP and its derivatives have now penetrated most biomedical research labs that use imaging as a tool. The importance of fluorescent probes in light microscopy can further be measured by the award of the Nobel Prize for Chemistry in 2008 to Tsien, Shimomura and Chalfie. The role of probes is as pertinent in the emerging field of correlative microscopy, though arguably far more complex. In correlative microscopy, two or more imaging modalities are applied to a single sample, with the combined images yielding more information than when each modality is used independently. To reflect this, we often use the phrase ‘1+1=3’. The most established correlative microscopy technique is correlative light electron microscopy (CLEM). Usually confocal LM and transmission EM (TEM) are correlated, though recently there have been major developments in both light and electron microscopy leading to other combinations. Correlative light and scanning EM in particular is a growing area due to the availability of new 3D automated microscopes like the serial
* Paul Verkade [email protected] 1
Lincoln’s Inn Fields Laboratory, The Francis Crick Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK
2
Wolfson Bioimaging Facility, Schools of Biochemistry and Physiology & Pharmacology, Medical Sciences Building, University of Bristol, University Walk, Bristol BS8 1TD, UK
block face SEM, the focused ion beam SEM, and array tomography [5]. However, whereas fluorescent molecules (either as an expression construct or through antibody or ligand coupling) are the most commonly used to detect a protein of interest in LM, this fluorescence is not directly visible in the electron microscope. Here, an electron-dense moiety such as a gold particle is generally required. Thus, the most direct way to produce a marker/probe (these terms are used interchangeably in the field and in this issue) is to couple both a fluorescent molecule and an electron-dense particle to the protein of interest. Unfortunately though, it is not that simple, and in most instances, the direct coupling of, e.g. a gold particle next to a fluorescent molecule decreases or completely masks the fluorescence emission (see, e.g. [2]). Hence, there have been major efforts to create the ‘optimal’ CLEM probe, one that is visible directly in the LM and EM and can be easily tagged to a protein of interest. Most likely there will never be a single optimal CLEM probe that can be used for all applications, and one has
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