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Measuring Diffusion Coefficients via Two-photon Fluorescence Recovery After Photobleaching
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Measuring Diffusion Coefficients via Two-photon Fluorescence Recovery After Photobleaching

Measuring Diffusion Coefficients via Two-photon Fluorescence Recovery After Photobleaching

DOI:
10.3791/1636-v

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07:00 min

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February 26, 2010

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,
1Department of Physics and Astronomy,University of Rochester, 2Department of Biomedical Engineering,University of Rochester

Chapters

  • 00:00Title
  • 00:46Introduction
  • 01:09Aligning the Optics
  • 02:51Determining Safe Monitor Powers
  • 03:54Determinging the Input Parameters to the Scaler and Pulse Generator
  • 06:00Representative Results
  • 06:31Conclusion

Summary

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In this article we will describe the procedure for measuring diffusion coefficients using multi-photon fluorescence recovery after photobleaching. We will begin by aligning the laser along the optical path to the sample and determining the proper experimental parameters, then continue generating and finally fitting fluorescence recovery curves.

Tags

Diffusion CoefficientsTwo-photon Fluorescence Recovery After PhotobleachingMulti-fluorescence Recovery After PhotobleachingMicroscopy TechniqueMacromoleculesIn VitroIn VivoBiological SystemsPhotobleachingLaser FlashFluorescence MonitoringPockels CellLaser ModulatorOptical PathSample PreparationAqueous Fluorescent DyeExperimental ParametersMonitor PowerBleaching PowerBleach DurationBin WidthsFluorescence Recovery TimeRecovery CurvesLabVIEW AutomationEnhanced ThroughputMathematical Model Fitting Algorithm

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