Photochemical damage from chronic 458-nm laser exposures in an artificially pigmented hTERT-RPE1 cell line

Michael S. Foltz; Norris A. Whitlock; Larry E. Estlack; Manuel A. Figueroa; Robert J. Thomas; Benjamin A. Rockwell; Michael L. Denton

doi:10.1117/12.644690

1 March 2006 Photochemical damage from chronic 458-nm laser exposures in an artificially pigmented hTERT-RPE1 cell line

Michael S. Foltz, Norris A. Whitlock, Larry E. Estlack, Manuel A. Figueroa, Robert J. Thomas, Benjamin A. Rockwell, Michael L. Denton

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Proceedings Volume 6084, Optical Interactions with Tissue and Cells XVII; 608415 (2006) https://doi.org/10.1117/12.644690
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

Artificially pigmented hTERT-RPE1 cells were exposed to a mode-locked or continuous wave (CW) laser at 458 nm for one hour in a modified culture incubator. Exposure conditions were selected to give greatest likelihood of damage due to a photochemical mechanism, with interest in possible differences between CW and mode-locked damage thresholds. After post-exposure-recovery (PER) for either 1-hour or 24-hour, cells were concurrently stained with annexin V and 6-CFDA to determine if they had undergone necrosis or apoptosis. Alternatively, cells were stained with Ethidium Homodimer (EthD-1) and Calcein AM to determine if they had undergone necrosis following 1-hour and 24-hours PER. Preliminary results indicate that laser exposure induced some apoptosis following 1-hour PER, with irradiance required for apoptosis being lower than that for necrosis with mode-locked exposure conditions. Probit analysis yielded necrosis thresholds for cell culture following 1-hour PER using data compiled from both dye sets. CW exposures resulted in a lower threshold than mode-locked exposures for necrosis following 1-hour PER. A thermal model provided the predicted temperature rise in cell culture due to laser exposure. The thermal model validates our choice of laser parameters to obtain photochemical damage. Data following 24-hours PER were inconclusive. Considerations of cell migration are included in the interpretation of data and further improvements to methods when using live cell assays are recommended.

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Michael S. Foltz, Norris A. Whitlock, Larry E. Estlack, Manuel A. Figueroa, Robert J. Thomas, Benjamin A. Rockwell, and Michael L. Denton "Photochemical damage from chronic 458-nm laser exposures in an artificially pigmented hTERT-RPE1 cell line", Proc. SPIE 6084, Optical Interactions with Tissue and Cells XVII, 608415 (1 March 2006); https://doi.org/10.1117/12.644690

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