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BP: Fachverband Biologische Physik

BP 17: Bioimaging

BP 17.9: Talk

Wednesday, March 20, 2024, 12:00–12:15, H 2032

CRISPR screen to improve the optical properties of living tissues — •Susan Wagner, Venkat R. Krishnaswamy, Kaushikaram Subramanian, Heike Petzold, Benjamin Seelbinder, and Moritz Kreysing — Institute of Biological and Chemical Systems - Biological Information Processing, KIT, Karlsruhe

Optical microscopy has been massively advanced to deliver unprecedented resolution allowing discoveries down to the molecular level. Nevertheless, optical access of living biological samples by microscopes is usually restricted to the outer most surface owing to tissue-induced light scattering.

We successfully improved the optical properties of mammalian cells and found that evolved transparency frequently goes along with the reduction of nuclear granularity, while the gene expression profile reflects scattering properties of cells. To genetically clear living mammalian tissues, we are conducting a genome-wide CRISPR activation screen to find those genes which confer transparency.

As a next step, we are investigating how improved optical properties of individual cells influence the optical properties of 3D cell clusters, such as spheroids, using interspersed fluorescent microspheres to quantify imaging quality.

Understanding the full range of a tissue's optical plasticity will provide us with a broad toolkit, so that different genetic strategies can be applied depending on the specific nature of the various biological samples.

Keywords: tissue transparency; living biological sample; microscopy; CRISPR screen