Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Nov;284(2):97-102.
doi: 10.1111/jmi.13056. Epub 2021 Sep 20.

Correlative light and volume electron microscopy (vCLEM): How community participation can advance developing technologies

Christopher J Guerin  1 , Saskia Lippens  1
Affiliations

Correlative light and volume electron microscopy (vCLEM): How community participation can advance developing technologies

Christopher J Guerin et al. J Microsc. 2021 Nov.

Abstract

Correlative light and electron microscopy is a valuable tool to image samples across resolution scales and link data on structure and function. While studies using this technique have been available since the 1960s, recent developments have enabled applying these workflows to large volumes of cells and tissues. Much of the development in this area has been facilitated through the collaborative efforts of microscopists and commercial companies to bring the methods, hardware and image processing technologies needed into laboratories and core imaging facilities. This is a prime example of how what was once a niche area can be brought into the mainstream of microscopy by the efforts of imaging pioneers who push the boundaries of possibility.

Keywords: correlative light and electron microscopy; correlative volume electron microscopy; electron microscopy; volume electron microscopy.

PubMed Disclaimer

Figures

FIGURE 1
FIGURE 1
Possible workflows for vCLEM. A number of light microscopic imaging techniques can be incorporated into a vCLEM experiment with the aim of identifying particular structures of interest usually by fluorescently staining a protein or structure of interest. Intermediate imaging steps using X‐Ray microscopy or micro CT can further assist in identifying a region of interest (ROI) to be targeted in the electron microscope. Typical vEM methods used would be in chamber sectioning either with a diamond knife (SBF‐SEM) or a focused ion beam (FIB‐SEM) or external sectioning of a resin block followed by serial imaging (AT) of the resulting sections. Supplemental imaging techniques such as EDX or mass spectrometry can be used to provide elemental or protein signatures. Finally, the LM and vEM images and any associated data are compiled in the registered sections and visualisation is carried out via computer modelling of the data stack
See this image and copyright information in PMC

References

    1. Linberg, K. A. , & Fisher, S. K . (1986). An ultrastructural study of interplexiform cell synapses in the human retina. Journal of Comparative Neurology, 243(4), 561–576. - PubMed
    1. Guerin, C. J. , Nalan, L. , & Klumperman, J . (2020). It's a small, small world: A brief history of biological correlative microscopy. In Verkade P. & Collinson L.M. (Eds.), Correlative imaging: Focusing on the future. Wiley: UK.
    1. <number>3.</number (1987). Correlative microscopy in biology: Instrumentation and methods. Hyatt (Ed.) M.A.. Orlando, Florida: Academic Press Inc.
    1. Hoppe, W. , et al. (1974). Three‐dimensional reconstruction of individual negatively stained yeast fatty‐acid synthetase molecules from tilt series in the electron microscope. Hoppe‐Seyler's Zeitschrift für Physiologische Chemie, 355(11), 1483–1487. - PubMed
    1. Young, R. J. , et al. (1993). An application of scanned focused ion beam milling to studies on the internal morphology of small arthropods. Journal of Microscopy, 172(1), 81–88.