Dresden 2014 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 13: Posters: DNA/RNA and related enzymes
BP 13.5: Poster
Dienstag, 1. April 2014, 09:30–12:30, P1
Single Molecule Localization Microscopy of Chromatin Structures — •Udo Birk1,2, Kirti Prakash1, Aleksander Szczurek1, Hyun-Keun Lee1, and Christoph Cremer1,2 — 1Institute of Molecular Biology (IMB), Ackermannweg 4, 55118 Mainz, Germany — 2Kirchhoff Institute for Physics, Heidelberg University, 69120 Heidelberg, Germany
Understanding the structural and organizational aspects of chromatin at different stages of cell differentiation as well as at various phases of the cell cycle is one of the many promising applications of superresolution microscopy. It is challenging to study e.g. the organization of the different proteins which form the basis of chromosomal superstructures, due to limitations of conventional Light Microscopy (LM) and of Electron Microscopy (EM). To this end, structured illumination microscopy (with an optical resolution of about 100 nm in the object plane) could provide an improved resolution of intact cell nuclei and of the chromatin therein.
We report results on visible light based single molecule localization microscopy (SMLM) of chromatin structures in intact cell nuclei, and studied these structures at different stages of the cell cycle using the SMLM method of Spectral Position Determination Microscopy (SPDM). We analyzed the distribution of DNA and of associated proteins. The images obtained show a dramatic increase in light optical resolution of chromatin structures. Novel labeling techniques are required to make full use of SMLM visualization of DNA also directly labeled, i.e. without fluorescence in-situ hybridization.