SKM 2023 – scientific programme

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BP: Fachverband Biologische Physik

BP 28: Poster Session II

BP 28.12: Poster

Thursday, March 30, 2023, 18:00–20:00, P2/EG

Screening for Pharmacologically Enhanced Tissues for Optical Microscopy. — •Venkat Raghavan Krishnaswamy, Susan Wagner, Rico Barsacchi, and Moritz Kreysing — Max Planck Institute of Molecular Cell Biology and Genetics. Dresden. Germany.

Our ability to observe the complexity of a living tissue under a light microscope is heavily impeded by the inherent light-scattering properties of the biological tissues. But remarkably some living tissues are highly transparent in nature, e.g., human retina, certain deep-sea creatures, etc. Comprehending and replicating the mechanisms by which tissue transparency is attained would unleash the full capabilities of optical microscopes. Here, we propose to unravel the molecular basis of tissue transparency in three-dimensional tissues using the immense potential of high-throughput screening and large-content image analysis tools. Specifically, we used active pharmacological compounds to screen for pathways that impact light-scattering properties in spheroids of human colorectal carcinoma cell line (HCT116). Randomly integrated multifluorescent beads within the spheroids served as an extrinsic readout factor and a conventional chemical-based clearing method was used as a positive control for assessing the optical properties of the tissue. The image analysis pipeline constructed to measure the intensities of the beads through the Z-sections clearly distinguishes cleared from uncleared spheroids. Overall, a robust assay was developed which would be employed on larger screens, to rapidly identify compounds and, further, signaling pathways that regulate the molecular mechanisms of tissue transparency.