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Review
. 2014 Oct 1;70:12.36.1-10.
doi: 10.1002/0471142956.cy1236s70.

Correlative Fluorescence and Electron Microscopy

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Free PMC article
Review

Correlative Fluorescence and Electron Microscopy

Randall T Schirra Jr et al. Curr Protoc Cytom. .
Free PMC article

Abstract

Correlative fluorescence and electron microscopy (CFEM) is a multimodal technique that combines dynamic and localization information from fluorescence methods with ultrastructural data from electron microscopy, to give new information about how cellular components change relative to the spatiotemporal dynamics within their environment. In this review, we will discuss some of the basic techniques and tools of the trade for utilizing this attractive research method, which is becoming a very powerful tool for biology labs. The information obtained from correlative methods has proven to be invaluable in creating consensus between the two types of microscopy, extending the capability of each, and cutting the time and expense associated with using each method separately for comparative analysis. The realization of the advantages of these methods in cell biology has led to rapid improvement in the protocols and has ushered in a new generation of instruments to reach the next level of correlation--integration.

Keywords: cell biology; correlative; cryoEM; electron microscopy; fluorescence; imaging; immunolabel; integrated microscope; light microscopy; tomography.

Figures

Figure 1
Figure 1
A general overview of methods that are to be considered when approaching a sample for ultrastructural localization and analysis in CFEM. First, the type of information required (localization vs spatiotemporal data) will determine the type of FM that is necessary and will lead to a decision about the best type of label and EM procedure to gain an overall picture of a protein or process using a correlated technique.
Figure 2
Figure 2
Flow chart for live cell correlation in CFEM. Samples are grown on EM grids (A) and the cultures are imaged in FM (B) prior to being fixed or vitrified and imaged in an EM with correlation (F) or vitrified (C) and reimaged on a cryo-FM (D) for subsequent imaging in EM and correlation (F).

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