Heidelberg 2015 – scientific programme
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MO: Fachverband Molekülphysik
MO 21: Posters 4: Novelties in Molecular Physics
MO 21.10: Poster
Thursday, March 26, 2015, 17:00–19:00, C/Foyer
Single Molecule Localisation Microscopy of the Distribution of Chromatin Nanostructures Using Hoechst and Dapi Fluorescent Probes — •Udo Birk1,2,3, Aleksander Szczurek1, Kirti Prakash1, Hyun-Keun Lee1,3, Dominika Zurek-Biesiada4, Gerrit Best2, Martin Hagmann2, Jurek Dobrucki4, and Christoph Cremer1,2,3 — 1Institute of Molecular Biology, Mainz, Germany — 2Heidelberg University, Germany — 3Institut für Physik, Mainz University, Germany — 4Jagiellonian University, Kraków, Poland
We present a Single Molecule Localization Microscopy application of specific fluorescent DNA dyes such as DAPI, Hoechst and others, in order to obtain chromatin/nuclear DNA density maps with high optical and structural resolution. The approach adopted here 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. In mammalian cell nuclei, this technique yielded a single molecule localization precision in the order of 15 - 30 nm, corresponding to an optical (two-point) resolution of roughly 40 - 70 nm. We investigated various DNA structures and obtained data with a DNA-bound fluorophore density as high as 5000 SM/micrometer^2, significantly higher than previously reported.
[1] A. Szczurek et al. Nucleus 2014.