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Humboldt-Universität zu Berlin - IRI Life Sciences

Humboldt-Universität zu Berlin | IRI Life Sciences | Events | Tutorials | Tutorial in Life Sciences: Yannick Schwab "CLEM"

Tutorial in Life Sciences: Yannick Schwab "CLEM"

When Sep 14, 2017 from 09:00 AM to 01:00 PM (Europe/Vienna / UTC200) iCal
Where IRI Life Sciences, Philippstr. 13, Bld. 18, 3rd Floor
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No registration required!

 

The Tutorial will be in two parts:

Lecture #1: Correlative Light and Electron Microscopy (CLEM)
CLEM regroups a set of techniques that aim to image the same region of a sample with both light and electron microscopy. The goal is to benefit from the advantages of both modalities. On one side, light microscopes, and more specifically fluorescence imaging benefit from a large field of view, can identify molecular species providing specific probes are utilized, and are compatible with imaging living samples. On the other side, electron microscopy uniquely reveals the rich and complex ultrastructure of the cells at high resolution.
This lecture will provide an overview of the methods currently used in CLEM and will cover practical aspects including sample preparation, probes, correlation strategies and dedicated tools applied to a variety of specimens from cells in culture to model organisms.
 
Lecture #2: Towards a quantitative description of subcellular phenotypes
by Correlative Light and Electron Microscopy 
A variety of strategies are now available to correlate live cell or in vivo imaging to EM, but they very often suffer from tedious workflows. The Schwab team activities are centred on the development of new methods that increase the throughput of correlation on both cultured cells and multicellular specimens. By elaborating on the capacities of the focused ion beam - scanning electron microscope, automated correlation enables the acquisition of dozens of selected cells in a fully unattended fashion. The selection is performed by high-throughput screening at the light microscopic level which defines objective criteria to select the sub-populations of cells to be further analysed by electron microscopy. As a result, impact of various treatments, such as siRNA, on organelles morphology and distribution can be analysed at the ultrastructural level on multiple cells.
On multicellular specimens, the major challenge is the navigation within the volume of the samples to reach to region of interest. Precise targeting is mandatory in order to make the correlation efficient. By using microscopic X-Ray computed tomography as a bridging modality, the 3D data obtained by in-vivo imaging, that reveal the targeted cells position, is registered to the topology of the resin embedded specimen. With this information, exposing the sub-volume of interest is fast and precise. This workflow opens to the transversal observation of dynamic events such a tumor-cell extravasation in a mouse model of brain metastasis.