A novel optimized pre-embedding antibody-labelling correlative light electron microscopy technique

Author:

Doyle Nicole1ORCID,Simpson Jennifer1,Hawes Philippa C.21ORCID,Maier Helena J.1ORCID

Affiliation:

1. The Pirbright Institute, Ash Road, Pirbright, Woking, GU24 0NF, UK

2. Present address: The Francis Crick Institute, Midland Road, London, NW1 1AT, UK

Abstract

In the intricate environment of a cell, many studies seek to discover the location of specific events or objects of interest. Advances in microscopy in recent years have allowed for high detail views of specific areas of cells of interest using correlative light electron microscopy (CLEM). While this powerful technique allows for the correlation of a specific area of fluorescence on a confocal microscope with that same area in an electron microscope, it is most often used to study tagged proteins of interest. This method adapts the correlative method for use with antibody labelling. We have shown that some cellular structures are more sensitive than others to this process and that this can be a useful technique for laboratories where tagged proteins or viruses, or dedicated CLEM instruments are not available.

Funder

Biotechnology and Biological Sciences Research Council

Publisher

Microbiology Society

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