Integrated light and electron microscopy

Book chapter (2019)

Authors

R. I. Koning Leiden University Medical Center
A. Srinivasa Raja ImPhys/Microscopy Instrumentation & Techniques - AS
R. Lane ImPhys/Microscopy Instrumentation & Techniques - AS
A. J. Koster Leiden University Medical Center
J.P. Hoogenboom ImPhys/Microscopy Instrumentation & Techniques - AS
Research Group
ImPhys/Microscopy Instrumentation & Techniques (AS) (TU Delft)
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Published Date

2019

Language

English

Reuse Rights

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Faculty

Applied Sciences

Department

ImPhys/Imaging Physics

Research Group

ImPhys/Microscopy Instrumentation & Techniques

Abstract

In the past decades, correlative light and electron microscopy (CLEM) methods have evolved from being mostly used by a few pioneering, specialist labs to a collection of techniques and workflows practiced by a broad group of researchers in structural biology. In most cases, CLEM involves a distinct set of sequentially used specimen preparation and labeling techniques, followed by diverse types of light and electron microscopy techniques. This chapter focuses on those areas in present-day CLEM that are faced with challenges for which these advantages of integrated microscopes may well be key for further advancement. These areas are large-scale and high-throughput correlated (volume) microscopy, super-resolution localization in resin or cryo-frozen sections, fluorescence-guided focused ion beam milling for cryo-electron tomography, and the integration of sample preparation and transfer. Ultimately this should lead to the development of specific integrated CLEM systems with complete and fully automated workflows, leading to high-throughput and high-yield systems.