Boudreau, Colton, Wee, Tse-Luen Erika, Duh, Yan-Rung Silvia, Couto, Melissa P, Ardakani, Kimya H, Brown, Claire M (August 2016) Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity. Scientific Reports, 6 (1). p. 30892. ISSN 2045-2322
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Abstract
It is important to determine the most effective method of delivering light onto a specimen for minimal light induced damage. Assays are presented to measure photo-bleaching of fluorophores and photo-toxicity to living cells under different illumination conditions. Turning the light off during part of the experimental time reduced photo-bleaching in a manner proportional to the time of light exposure. The rate of photo-bleaching of EGFP was reduced by 9-fold with light pulsing on the micro-second scale. Similarly, in living cells, rapid line scanning resulted in reduced cell stress as measured by mitochondrial potential, rapid cell protrusion and reduced cell retraction. This was achieved on a commercial confocal laser scanning microscope, without any compromise in image quality, by using rapid laser scan settings and line averaging. Therefore this technique can be implemented broadly without any software or hardware upgrades. Researchers can use the rapid line scanning option to immediately improve image quality on fixed samples, reduce photo-bleaching for large high resolution 3D datasets and improve cell health in live cell experiments. The assays developed here can be applied to other microscopy platforms to measure and optimize light delivery for minimal sample damage and photo-toxicity.
| Item Type: | Paper |
|---|---|
| Subjects: | Investigative techniques and equipment Investigative techniques and equipment > microscopy |
| CSHL Authors: | |
| Communities: | CSHL labs > Spector lab |
| SWORD Depositor: | CSHL Elements |
| Depositing User: | CSHL Elements |
| Date: | 3 August 2016 |
| Date Deposited: | 04 Jan 2024 20:30 |
| Last Modified: | 04 Jan 2024 20:30 |
| PMCID: | PMC4971503 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/41381 |
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