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SEMINAR: CMCA Seminar: Super-resolution optical imaging of chromatin, DNA damage and repair

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Today's date is Tuesday, December 01, 2020
CMCA Seminar: Super-resolution optical imaging of chromatin, DNA damage and repair : A new approach to imaging chromatin in situ Other events...
Critical aspects of structure and function of the cell nucleus are often inaccessible to wide field and confocal imaging. Higher order chromatin structures, subnuclear bodies, repair foci etc., cannot be imaged in detail without applying 'super-resolution' techniques. A new approach to imaging chromatin in situ has become possible by exploiting photoconversion of UV-excited DNA dyes Hoechst 33258, DAPI, and Vybrant® DyeCycle™ Violet. Single Molecule Localization Microscopy (SMLM) is based on using two wavelengths of light - one for regeneration of the pool of the blinking form of the dye, and the other for excitation, or just one high intensity excitation light to transfer the dye between the emitting and non-emitting states. SMLM enables optical isolation and localization of high numbers of DNA-bound molecules, usually in excess of 106 in one cell nucleus. This approach yields images of DNA density with the resolution several times better than conventional optical microscopy, reaching 40 - 50 nm in the specimen plane, and offers several important advantages over the previously described imaging methods, including an ability to record images using a single wavelength excitation of a relatively low intensity, and a higher density of single molecule signals than in previous studies. High resolution images (SMLM, dSTORM, SIM) of chromatin based on phototonversion of UV-excited DNA dyes were combined with images representing scheduled and unscheduled DNA replication (EdU, click reaction), histone H2AX phosphorylation (marking DNA double strand breaks), and XRCC1 repair factor (single strand breaks) in order to study the mechanisms of induction of DNA damage and repair.
Speaker(s) Professor Jurek Dobrucki, Jagiellonian University in Kraków
Location CMCA Seminar Room 1.80, Physics Building
Contact CMCA Admin <[email protected]> : 6488 5387
Start Thu, 30 Oct 2014 16:00
End Thu, 30 Oct 2014 17:00
Submitted by CMCA Admin <[email protected]>
Last Updated Tue, 28 Oct 2014 09:32
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