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BP: Fachverband Biologische Physik

BP 24: Physics of Cells II

BP 24.6: Vortrag

Donnerstag, 17. März 2011, 11:45–12:00, ZEU 250

Using novel microscopy methods to correlate pluripotent stem cell state with subcellular structure — •Kevin Chalut, Markus Hoepfler, Andrew Ekpenyong, and Jochen Guck — Cavendish Laboratory, University of Cambridge, Cambridge, UK

The function of pluripotent stem cells (PSCs) is to commit to all types of tissue cells needed for an organism while self-renewing and maintaining their pluripotency until all lineages are established. PSC state - pluripotent, pre-committed, or committed - has primarily been probed by investigating biochemical properties, but the mystery of how biological diversity is established while maintaining pluripotency remains unsolved. In an effort to solve this mystery, we probed PSC state by evaluating their physical properties. These physical properties include their internal structure, particularly changes in chromatin structure. To visualise the relationship between chromatin structure and PSC state, we used a fluorescent label for heterochromatin proteins, and then imaged using confocal microscopy and STED. Furthermore, we used digital holographic microscopy, a live-cell and label-free technique, to visualise chromatin structure and correlate it with PSC state. We saw in all techniques that, prior to differentiation, the chromatin structure opens up considerably, diffusing throughout the nucleus. This opening up of chromatin correlates with greater transcriptional accessibility. These structural changes are a physical phenotype that we can use to deduce PSC state, and they can also be used as a biomarker for pluripotency and differentiation.