Confocal Microscopy for Laser-Induced Damage

The Olympus FV/1000 confocal microscopy system (Olympus) and FV/1000 software were used for image acquisition and analysis and described in a previous study (22 (link)). Damage was induced with a 405 nm laser. The laser passed through a PLAON 60X oil lens. The output power was 5 mW/scan. Cells transfected with GFP-tagged proteins were incubated at 37°C on a thermos plate in normal media during observation. For the evaluation of accumulation and kinetics, the mean intensity of each accumulated point or line was obtained after subtraction and quantified by ImageJ.

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Gao Y., Li C., Wei L., Teng Y., Nakajima S., Chen X., Xu J., Legar B., Ma H., Spagnol S.T., Wan Y., Dahl K.N., Liu Y., Levine A.S, & Lan L. (2017). SSRP1 cooperates with PARP and XRCC1 to facilitate single strand DNA break repair by chromatin priming. Cancer research, 77(10), 2674-2685.

Publication 2017

FV/1000 confocal microscopy system FV/1000 software

Cells Confocal microscopy Kinetics Lens Proteins Scan

Corresponding Organization : University of Pittsburgh

Other organizations : Tsinghua University, General Hospital of Shenyang Military Region, Carnegie Mellon University

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Variable analysis

independent variables

Laser wavelength (405 nm)
Laser power (5 mW/scan)

dependent variables

Accumulation and kinetics of GFP-tagged proteins
Mean intensity of accumulated points or lines

control variables

Cell culture conditions (incubated at 37°C on a thermos plate in normal media during observation)
Imaging system (Olympus FV/1000 confocal microscopy system and FV/1000 software)

controls

Positive control: Cells transfected with GFP-tagged proteins
Negative control: Not specified

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