Dataset Information

Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

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.

SUBMITTER: Boudreau C

PROVIDER: S-EPMC4971503 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature

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Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

Boudreau Colton C Wee Tse-Luen Erika TL Duh Yan-Rung Silvia YR Couto Melissa P MP Ardakani Kimya H KH Brown Claire M CM

Scientific reports 20160803

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 ...[more]

PMID: 27485088

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Project description:AimThe study aimed to compare teeth sensitivity and shade after bleaching protocol with descending different light intensities versus bleaching protocol with the same high light intensity.Material and methodsSample size was twenty-four patients. Each group consisted of twelve patients. Group, I patients received bleaching protocol of descending different light intensities. Group II patients received bleaching protocol with the same high light intensity; both groups used the same home bleaching gel kit for seven days according to manufacturer instructions and protocol. Baseline records were digital photographs, teeth sensitivity and teeth shade for 12 anterior teeth. Teeth sensitivity was assessed using five points verbal rating scale and Standardized 100 mm Visual analogue scale after 1 day, after 2 days and after 1 week. Teeth shades for twelve anterior teeth were recorded by VITA Easy Shade V (VITA Zahnfabrik H. Rauter GmbH & Co. KG, Germany) after 1 week by VITA Easy Shade V. Mann-Whitney test (non-parametric test, 2 independent samples) was used to compare teeth sensitivity between both bleaching protocols at each period. A paired t-test (parametric test, 2 related samples) was performed to compare the colour change in shade guide units (SGU) and ∆E values within high light intensity bleaching protocol. While Wilcoxon Signed-Rank test (non-parametric test, 2 related samples) was used to compare colour change light intensities bleaching protocol. Comparison of bleaching effectiveness (∆SGU and ∆Ediff) between both bleaching protocols was performed by the Mann-Whitney test.ResultsDescending light intensities protocol showed a lower teeth sensitivity than high light intensity protocol after 1 and 2 days. There was no teeth sensitivity reported at 1-week post-bleaching. Regarding the teeth shade, descending light intensities protocol had a little higher effect on colour change in shade guide units (SGU) than high light intensity protocol effect. Both bleaching protocols showed there was no significant difference in ∆SGU recorded after bleaching between high and descending light intensities protocols.ConclusionDescending different light intensities protocol showed a lower teeth sensitivity than high same light intensity protocol. Descending light intensities protocol had a little higher effect on colour change in shade guide units (SGU) than high light intensity protocol effect.

Project description:BACKGROUND:At the forefront of ecosystems adversely affected by climate change, coral reefs are sensitive to anomalously high temperatures which disassociate (bleaching) photosynthetic symbionts (Symbiodinium) from coral hosts and cause increasingly frequent and severe mass mortality events. Susceptibility to bleaching and mortality is variable among corals, and is determined by unknown proportions of environmental history and the synergy of Symbiodinium- and coral-specific properties. Symbiodinium live within host tissues overlaying the coral skeleton, which increases light availability through multiple light-scattering, forming one of the most efficient biological collectors of solar radiation. Light-transport in the upper ~200 ?m layer of corals skeletons (measured as 'microscopic' reduced-scattering coefficient, ?'(S,m)), has been identified as a determinant of excess light increase during bleaching and is therefore a potential determinant of the differential rate and severity of bleaching response among coral species. RESULTS:Here we experimentally demonstrate (in ten coral species) that, under thermal stress alone or combined thermal and light stress, low-?'(S,m) corals bleach at higher rate and severity than high-?'(S,m) corals and the Symbiodinium associated with low-?'(S,m) corals experience twice the decrease in photochemical efficiency. We further modelled the light absorbed by Symbiodinium due to skeletal-scattering and show that the estimated skeleton-dependent light absorbed by Symbiodinium (per unit of photosynthetic pigment) and the temporal rate of increase in absorbed light during bleaching are several fold higher in low-?'(S,m) corals. CONCLUSIONS:While symbionts associated with low-[Formula: see text] corals receive less total light from the skeleton, they experience a higher rate of light increase once bleaching is initiated and absorbing bodies are lost; further precipitating the bleaching response. Because microscopic skeletal light-scattering is a robust predictor of light-dependent bleaching among the corals assessed here, this work establishes ?'(S,m) as one of the key determinants of differential bleaching response.

Dataset Information

Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

Publications

Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

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