Elsevier

Methods in Cell Biology

Volume 111, 2012, Pages xvii-xix
Methods in Cell Biology

Preface

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Introduction to Correlative Light and Electron Microscopy

Correlative microscopy, as an approach to combine methods of different modalities, has become increasingly important over the past years. With the advent of GFP and the accompanying improvements in light as well as in electron microscopy technology, it is the combination of both techniques in Correlative Light and Electron Microscopy (CLEM) that has generated the most attention. The rise in the importance of CLEM is reflected by a steady increase in publications related to the fusion of these

Acknowledgments

The authors would like to thank Shaun Gamble (Elsevier) for his help in bringing this volume of Methods in Cell Biology to completion.

References (13)

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Cited by (10)

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    Correlative light and electron microscopy (CLEM) is a multimodal tool that enables the imaging of the same specimen by both light and electron microscopy (Müller-Reichert & Verkade, 2012).

  • HPM live μ for a full CLEM workflow

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    However, EM can provide full contextual information down to sub-nanometer details, although specialized techniques (e.g. the Tokuyasu technique (Geuze, Slot, Van Der Ley, & Scheffer, 1981)) may be required to identify specific objects. With a well-designed sample preparation/imaging workflow for CLEM (Mironov, Polishchuk, & Luini, 2000; Polishchuk, Polishchuk, & Luini, 2012) —where LM is typically used prior to EM we can preserve the benefits of the two techniques (de Boer, Hoogenboom, & Giepmans, 2015; Heiligenstein, Paul-Gilloteaux, Raposo, & Salamero, 2017; Müller-Reichert & Verkade, 2017; Müller-Reichert et al., 2014; Müller-Reichert & Verkade, 2012; Plitzko, Rigort, & Leis, 2009). This is particularly the case when high pressure freezing (HPF) is used, allowing direct cryofixation of biological samples without any chemical fixation (Akiva et al., 2015, 2019).

  • Cryo-fluorescence microscopy of high-pressure frozen C. elegans enables correlative FIB-SEM imaging of targeted embryonic stages in the intact worm

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    Conventional protocols involve chemical fixation during sample preparation for either light microscopy (LM) or electron microscopy (EM) or both; the accompanying correlation may be for spatial identification of the region of interest (ROI) by LM and EM, or for colocalization of specific probes. For an in-depth CLEM primer, the reader is directed to a previous volume of this publication and the references therein (Muller-Reichert & Verkade, 2012). Reliable cryogenic fixation and technological innovations have significantly advanced cryo-imaging of thin cellular specimens.

  • Observation and characterisation of macrophages in zebrafish liver

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    Structural and functional similarities were observed and compared to Kupffer cells in the human and rodent livers (for reviews, see Jones and Summerfield, 1988; Wisse et al., 1996). We applied correlative light and electron microscopy (CLEM), a concept effective in cell image analysis by localising, identifying and studying the functional and structural properties of the cells' of interest across different imaging platforms (Kobayashi et al., 2012; Müller-Reichert and Verkade, 2012). Noteworthy, previous correlated fluorescence-, and scanning electron microscopy (SEM) studies in zebrafish have already demonstrated the advantageous nature of this approach in disclosing cellular transport and distribution of albumin in the zebrafish liver (Cheng et al., 2019) including the localisation of cellular targets within large tissue volumes when combined with serial sectioning techniques (Wacker et al., 2016).

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