Analysis of cytoplasmic diffusion with multiphoton microscopy
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 vol...
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| Format | Conference Proceeding |
| Language | English |
| Published |
SPIE
09.05.2002
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| Online Access | Get more information |
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| Summary: | 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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| Bibliography: | Conference Date: 2002-05-09|2002-05-10 Conference Location: Ottawa, Ontario, Canada |
| ISSN: | 0277-786X |
| DOI: | 10.1117/12.2283911 |