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BP: Fachverband Biologische Physik
BP 20: Poster VIII
BP 20.1: Poster
Dienstag, 17. März 2020, 14:00–16:00, P2/4OG
High-throughput scanning SAXS of desmin-expressing cells — •Chiara Cassini1, Manfred Burghammer2, Harald Herrmann3, and Sarah Köster1 — 1IRP, Georg-August-Universität Göttingen, Germany — 2ESRF, Grenoble, France — 3DKFZ, Heidelberg, Germany
Desmin is the main intermediate filament (IF) protein in muscle cells. Recently, a large number of mutations in the desmin gene have been discovered to be pathogenic. In order to assess the structures formed in cells by normal and mutant desmin, a high resolution method, capable of retrieving structural information at sub-cellular length scales, without the need for slicing the cells, is preferable. Thus, we used scanning small angle X-ray scattering (SAXS) on three different cell lines generated from IF-free mouse fibroblasts: one expressing wild type desmin, one expressing R406W-desmin, and the IF-free mother cell clone itself. The cells were grown on silicon nitride windows and measured in freeze-dried state. Each window contained tens to hundreds of cells. Each cell scan used to take minutes to hours; recently, we were able to employ a special fast scanning mode that allowed us to image an entire window in about 8 hours only. This approach ensured the collection of a statistically significant pool of data in a reasonable time span. The large quantity of data thus collected was treated with a combination of semi-automated segmentation of the dark field images and parallel computations. In the end, we were able to carry out a statistically relevant comparison of local structure-related parameters of the three cell lines, such as anisotropy and orientation.