Some Guiding Principles for a "Simple" Correlative Light Electron Microscopy Experiment

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2024 May 6

PMID 38709480

Authors

Elina Mantyla

Paul Verkade

Affiliations

Soon will be listed here.

Abstract

In recent years, Correlative Multimodal Imaging (CMI) has become an "en vogue" technique and a bit of a buzzword. It entails combining information from different imaging modalities to extract more information from a sample that would otherwise not be possible from each individual technique. The best established CMI technology is correlative light and electron microscopy (CLEM), which applies light and electron microscopy on the exact same sample/structure. In general, it entails the detection of fluorescently tagged proteins or structures by light microscopy and subsequently their relative intracellular localization is determined with nanometer resolution using transmission electron microscopy (TEM). Here, we describe the different steps involved in a "simple" CLEM approach. We describe the overall workflow, instrumentation, and basic principles of sample preparation for a CLEM experiment exploiting stable expression of fluorescent proteins.

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