The use of a laser for correlating light and electron microscopic images in thick tissue specimens

Derron Bishop, Ivana Nikic, Martin Kerschensteiner, Thomas Misgeld

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

7 Scopus citations

Abstract

Correlating images between light and electron microscopy is difficult especially in tissue specimens with a substantial z-dimension. To facilitate correlated light and electron microscopy (CLEM) in thick tissue specimens, we describe a basic method of using a femto-pulsed near-infrared laser to burn precise three-dimensional fiducial markers that circumscribe cells or regions of interest for easy identification between imaging methods. This rapid and reliable approach permits traditional fixation and avoids the use of electron-dense labeling methods that can obscure ultrastructural details. The versatility of the technique permits CLEM in a variety of tissue specimens to allow interpretation of highly resolved ultrastructural data in the more macroscopic and potentially dynamic context of light microscopy.

Original languageEnglish
Title of host publicationMethods in Cell Biology
PublisherAcademic Press Inc.
Pages323-337
Number of pages15
DOIs
StatePublished - 2014

Publication series

NameMethods in Cell Biology
Volume124
ISSN (Print)0091-679X

Keywords

  • Confocal microscopy
  • Electron microscopy
  • In vivo
  • Multiphoton microscopy
  • Serial section reconstruction
  • Technique

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