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Phototoxicity is a universal problem in optical imaging. Effective and quantitative evaluation of phototoxicity allows for the optimization of imaging conditions and the minimization of functional perturbation to live biological samples. We found that microtubule polymerization dynamics is a sensitive and reliable readout that can quantify phototoxicity in various imaging conditions. We quantified the photosensitizing effect of common organelle stains under the illumination of different laser wavelengths and compared the phototoxicity of different conditions. By controlling oxygen levels during light treatment, we found that the type I photosensitizing effect is involved in the hypoxia condition. We also evaluated the photoperturbation of IR laser pulses at different average and peak power levels in nonlinear optical imaging. Applying a recently developed real-time precision opto-control system, phototoxicity and reactive oxygen species generated at specific organelles can be studied.
Chi Zhang,Shivam Mahapatra,Seohee Ma,Matthew Clark,Bin Dong, andMark Carlsen
"Evaluation of the phototoxicity in optical imaging using protein dynamics", Proc. SPIE PC12855, Advanced Chemical Microscopy for Life Science and Translational Medicine 2024, PC1285505 (13 March 2024); https://doi.org/10.1117/12.3006557
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Chi Zhang, Shivam Mahapatra, Seohee Ma, Matthew Clark, Bin Dong, Mark Carlsen, "Evaluation of the phototoxicity in optical imaging using protein dynamics," Proc. SPIE PC12855, Advanced Chemical Microscopy for Life Science and Translational Medicine 2024, PC1285505 (13 March 2024); https://doi.org/10.1117/12.3006557