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A Correlative Light and Electron Microscopy Method Based on Laser Micropatterning and Etching

  • Julien Colombelli
  • Carolina Tängemo
  • Uta Haselman
  • Claude Antony
  • Ernst H.K. Stelzer
  • Rainer Pepperkok
  • Emmanuel G. Reynaud
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 457)

Summary

Correlative microscopy is a hybrid method that allows the localization of events observed under visible, ultraviolet, or infrared light, at molecular and submolecular levels, combining two microscopy techniques. However, the main limitation of correlative microscopy is to develop a labeling technique that can be easily used first in light and then in electron microscopy. Laser etching is a well-established method to create precisely designed shapes or volumes in various materials including glass. We have applied this technique to develop a new correlative light and electron microscopy method and to apply it in our study of the Golgi apparatus. The location of the cell of interest is laser-inscribed into the glass allowing a simple follow-up in light and fluorescence microscopy. Furthermore, the glass surface is laser-etched and upon fixation and flat embedding, the inverse ridge can be localized as well as the cell of interest, which is then processed for electron microscopy.

Key Words

Correlative electron microscopy Golgi apparatus laser etching laser micropatterning light fluorescence microscopy plasma-induced ablation pulsed laser TEM 

Notes

Acknowledgments

Alfons Riedinger for software implementation of the laser surgery platform. This research was partially supported by the VDI-TZ and the German ministry for research and development (BMBF) by grant FKZ 13N8287.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Julien Colombelli
    • 1
  • Carolina Tängemo
    • 1
  • Uta Haselman
    • 1
  • Claude Antony
    • 1
  • Ernst H.K. Stelzer
    • 1
  • Rainer Pepperkok
    • 1
  • Emmanuel G. Reynaud
    • 1
  1. 1.Cell Biology and Cell Biophysics Unit, EMBLHeidelbergGermany

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