How to Successfully Perform Live-cell CLEM

Koenig, Julia

October 04, 2021

How to Successfully Perform Live-cell CLEM

Coral Nano

Coral Nano provides a streamlined live-cell CLEM solution for getting insight about structural changes of cellular components over time. Besides the technical handling described in the workflow manual, this article provides additional knowledge for a successful experimental run.

For this example, the main interest was the final separation of two mitotic cells, called abscission. After cytokinesis, the two dividing cells are only connected by an intercellular bridge which needs to be resolved. Because of its size, this mechanism can’t be fully understood by live-cell imaging. Therefore, the ultrastructural analysis is needed to understand the underlying mechanism of final cell separation.

Chapter overview

Preparation of sapphires
Seeding of cells
Live-cell imaging
Freeze substitution and embedding
Sample retrieval and sectioning
Data postprocessing
Image correlation

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