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Team : SIBARITA

Research Projects

Single-molecule-based High Content Screening

Project Leader(s): Jean-Baptiste Sibarita

Collaborations: D. Choquet, M. Sainlos, G. Giannone, M. Mondin (BIC), D. Bourgois (IBS), Sanofi.

We have developed an innovative high-content-screening platform allowing to quantify protein organization and dynamics, as well as fluorophore properties, at the single molecule level with HCS standards. This HCS-SMLM platform allows, in an automatic workflow to: i) screen an entire P96-wellplate in dSTORM, (spt)PALM or DNA-PAINT, ii) analyze on the fly tens of different quantitative descriptors, and iii) represent them in an intelligible manner using dedicated classification, data mining and representation tools (Beghin et al., Nature Methods, 2017).

We are developing an assay for drug screening and fluorescent molecule engineering. We are making a pilot experiment with the goal to functionally characterize the membrane and subcellular trafficking of several antibodies targeting receptors expressed at the surface of cells of interest.

While quantitative SMLM attracts lots of interest, it still performs relatively poorly. Central to this problem is the generally far-from-ideal efficiency of labeling procedures, both in terms of target specificity, preservation of biological function, and photophysical performance of the fluorescent markers. We aim to tackle this limit using HCS-SMLM to engineer better fluorescent proteins and optimize their working conditions (illuminations, buffers).

Fundings

None

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