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bioRxiv · 10.1101/2024.10.21.619370

Live-HPF CLEM using the HPM Live μ: Finding back all needles in every haystack. The last frontier in CLEM

Heiligenstein, X.·Kodera, C.·Bret, Y.·Muczynski, V.·Heiligenstein, J.·Belle, M.
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Abstract

Correlative light and electron microscopy is a scientific method that encompasses several technologies and workflows. One of the pioneering workflows consisted in vitrifying by high pressure freezing a sample, shortly after live observation1. Despite its extraordinary potential, it did not turn into a routine technique for practical reasons. We redesigned the entire tool set, from the sample carrier to the high-pressure freezing machine, to standardize and democratise the technique. In our manuscript, we present all the technological developments that lead to a routine workflow for live to HPF CLEM. We demonstrate our ability to track rapidly moving endosomes live and retrieve them at the electron microscopy level, in three dimensions, with high confidence.

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Heiligenstein, X., Kodera, C., Bret, Y., Muczynski, V., Heiligenstein, J., Belle, M.. 2024-10-21. Live-HPF CLEM using the HPM Live μ: Finding back all needles in every haystack. The last frontier in CLEM. https://doi.org/10.1101/2024.10.21.619370

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