4.4 Article

The CryoCapsule: Simplifying Correlative Light to Electron Microscopy

Journal

TRAFFIC
Volume 15, Issue 6, Pages 700-716

Publisher

WILEY
DOI: 10.1111/tra.12164

Keywords

correlative light to electron microscopy; CryoCapsule; endosomal network; freeze substitution; high-pressure freezing; image registration; langerin; melanosomes; spatial resolution; temporal resolution; Xenopus laevis mitotic spindle

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Funding

  1. EMBL PhD program
  2. ANR MatetPro Microconnect
  3. NIH [R01 EY015625]
  4. ARC [SL220100601359]
  5. CNRS
  6. Institut Curie
  7. French National Research Agency [ANR-10-INSB-04]

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Correlating complementary multiple scale images of the same object is a straightforward means to decipher biological processes. Light microscopy and electron microscopy are the most commonly used imaging techniques, yet despite their complementarity, the experimental procedures available to correlate them are technically complex. We designed and manufactured a new device adapted to many biological specimens, the CryoCapsule, that simplifies the multiple sample preparation steps, which at present separate live cell fluorescence imaging from contextual high-resolution electron microscopy, thus opening new strategies for full correlative light to electron microscopy. We tested the biological application of this highly optimized tool on three different specimens: the in vitro Xenopus laevis mitotic spindle, melanoma cells over-expressing YFP-langerin sequestered in organized membranous subcellular organelles and a pigmented melanocytic cell in which the endosomal system was labeled with internalized fluorescent transferrin.

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