Heidelberg 2015 – wissenschaftliches Programm
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Q: Fachverband Quantenoptik und Photonik
Q 62: Poster: Quantum Optics and Photonics III
Q 62.70: Poster
Donnerstag, 26. März 2015, 17:00–19:00, C/Foyer
Single molecule localization microscopy of chromatin structure using standard DNA dyes — Aleksander Szczurek1, •Christoph Cremer1,2,3, and Udo Birk1,3 — 1Institute of Molecular Biology, Mainz — 2Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg — 3Kirchhoff Institute for Physics, Heidelberg University, Heidelberg
In order to investigate DNA-chromatin structure, one may employ optical microscopy as a method of choice, due to feasibility. However, conventional light optical imaging approaches suffer from diffraction of the visible wavelengths used, limiting the resolution of structures to about 200 nm laterally and 600 nm axially. To overcome this shortcoming, recently novel visible light super-resolution imaging approaches have emerged. Presently, particulrly Structured Illumination Microscopy (SIM) and Single Molecule Localisation Microscopy (SMLM) have been established as useful approaches in investigations of eukaryotic cell nucleus. Here we present a novel application of commonly used DNA dyes in order to obtain nuclear DNA density maps with high optical and structural resolution. The approach presented is based on photoconversion to the green-emitting form of these dyes that later may undergo a process of switching under high intensity blue light [1][2]. In mammalian cell nuclei, this technique yielded a single molecule localisation precision in the order of 15 - 30 nm, corresponding to an optical resolution of roughly 40 - 70 nm.