29 August 2017 Analysis of cytoplasmic diffusion with multiphoton microscopy
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Proceedings Volume 10313, Opto-Canada: SPIE Regional Meeting on Optoelectronics, Photonics, and Imaging; 1031337 (2017) https://doi.org/10.1117/12.2283911
Event: Opto-Canada: SPIE Regional Meeting on Optoelectronics, Photonics, and Imaging, 2002, Ottawa, Ontario, Canada
Abstract
Fluorescence Photobleaching Recovery (FPR) has proven to be a versatile technique for assessing the transport phenomena of fluorescently tagged proteins. Effective diffusion coefficients are estimated by observing the rate of fluorescence recovery resulting from the influx of fluorophores into a volume rendered non-fluorescent by photobleaching. Diffusion coefficients may be determined through non-linear regression of a theoretical model' or by relating half times of recovery to calibration data2. Due to the lack of axial confinement inherent in confocal FPR, analyses are limited to two-dimensional systems. However, with the development of multiphoton microscopy, three-dimensional diffusion coefficients can now be determined with a spatial resolution of a few microns3.
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Eric J. Jervis, Eric J. Jervis, } "Analysis of cytoplasmic diffusion with multiphoton microscopy", Proc. SPIE 10313, Opto-Canada: SPIE Regional Meeting on Optoelectronics, Photonics, and Imaging, 1031337 (29 August 2017); doi: 10.1117/12.2283911; https://doi.org/10.1117/12.2283911
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