title: Retracing in correlative light electron microscopy: where is my object of interest?
abstract: Correlative light electron microscopy (CLEM) combines the strengths of light and electron microscopy in a single experiment. There are many ways to perform a CLEM experiment and a variety of microscopy modalities can be combined either on separate instruments or as completely integrated solutions. In general, however, a CLEM experiment can be divided into three parts: probes, processing, and analysis. Most of the existing technologies are focussed around the development and use of probes or describe processing methodologies that explain or circumvent some of the compromises that need to be made when performing both light and electron microscopy on the same sample. So far, relatively little attention has been paid to the analysis part of CLEM experiments. Although it is an essential part of each CLEM experiment, it is usually a cumbersome manual process. Here, we briefly discuss each of the three above-mentioned steps, with a focus on the analysis part. We will also introduce an automated registration algorithm that can be applied to the analysis stage to enable the accurate registration of LM and EM images. This facilitates tracing back the right cell/object seen in the light microscope in the EM.
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom.
authors.forename: Lorna
authors.initials: L
authors.lastname: Hodgson
authors.affiliation: Visual Information Laboratory, University of Bristol, Bristol, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Nam
authors.affiliation: Wolfson Bioimaging Facility, University of Bristol, Bristol, United Kingdom.
authors.forename: Judith
authors.initials: J
authors.lastname: Mantell
authors.affiliation: Visual Information Laboratory, University of Bristol, Bristol, United Kingdom.
authors.forename: Alin
authors.initials: A
authors.lastname: Achim
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom; Wolfson Bioimaging Facility, University of Bristol, Bristol, United Kingdom; School of Physiology & Pharmacology, University of Bristol, Bristol, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Verkade
mesh_terms: D000818:Animals; D055032:Electron Microscope Tomography / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence / Q000379:methods; D013194:Staining and Labeling
keywords: CLEM; Correlative light electron microscopy; Electron microscopy; Electron tomography; Overlay mapping; Retracing