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Photoconversion of Hoechst
Version 2 2019-04-08, 11:22
Version 1 2019-01-14, 16:57
dataset
posted on 2019-04-08, 11:22 authored by Verena HurstVerena Hurst, Susan GasserThe data set provided is a time series of image stacks showing the 405 nm laser-induced emission change of the DNA intercalating dye Hoechst from blue to green in living U2OS cells.
It is part of the following research note:
Hurst V and Gasser SM. The study of protein recruitment to laser-induced DNA lesions can be distorted byphotoconversion of the DNA binding dye Hoechst. F1000Research , :104.
It is part of the following research note:
Hurst V and Gasser SM. The study of protein recruitment to laser-induced DNA lesions can be distorted byphotoconversion of the DNA binding dye Hoechst. F1000Research , :104.
U2OS cells (a gift from Prof. Primo Leo Schaer, Department of Biomedicine, University of Basel) were incubated with 1.5 µg/ml Hoechst 33342 (Thermo Fisher Scientific, H1399) for at least 30 minutes prior to photoconversion. Photoconversion was induced with a VisiFRAP module (Visitron) mounted on the backport of the microscope and equipped with a 405 nm laser (Toptica, illumination power at the objective 12.8 mW). Confocal images were acquired with an Olympus IX81 microscope equipped with a PlanApo 100x/1.45 TIRFM oil objective, a CSU-X1 scan-head (Yokogawa), an Evolve 512 EMCCD camera (Photometrics), a 491nm laser (Cobolt Calypso 100), a 488/568 dichroic (Semrock Di01-T488/568-13x15x0.5), a band-pass 525/40 emission filter (Semrock FF01-525/40-25) and controlled with the Visiview Software (Visitron).
