Back to Search
Start Over
Characterizing observation volumes and the role of excitation saturation in one-photon fluorescence fluctuation spectroscopy
- Source :
- Journal of Biomedical Optics. 10:044015
- Publication Year :
- 2005
- Publisher :
- SPIE-Intl Soc Optical Eng, 2005.
-
Abstract
- Fluorescence correlation spectroscopy (FCS) and related distribution analysis techniques have become extremely important and widely used research tools for analyzing the dynamics, kinetics, interactions, and mobility of biomolecules. However, it is not widely recognized that photophysical dynamics can dramatically influence the calibration of fluctuation spectroscopy instrumentation. While the basic theories for fluctuation spectroscopy methods are well established, there have not been quantitative models to characterize the photophysical-induced variations observed in measured fluctuation spectroscopy data under varied excitation conditions. We introduce quantitative models to characterize how the fluorescence observation volumes in one-photon confocal microscopy are modified by excitation saturation as well as corresponding models for the effect of the volume changes in FCS. We introduce a simple curve fitting procedure to model the role of saturation in FCS measurements and demonstrate its accuracy in fitting measured correlation curves over a wide range of excitation conditions.
- Subjects :
- Materials science
Statistics as Topic
Biomedical Engineering
Fluorescence correlation spectroscopy
Models, Biological
Sensitivity and Specificity
Molecular physics
Fluorescence spectroscopy
Cell Physiological Phenomena
Biomaterials
Imaging, Three-Dimensional
Optics
Image Interpretation, Computer-Assisted
Microscopy
Computer Simulation
Spectroscopy
Cell Size
Photons
business.industry
Reproducibility of Results
Image Enhancement
Atomic and Molecular Physics, and Optics
Electronic, Optical and Magnetic Materials
Spectrometry, Fluorescence
Microscopy, Fluorescence
Curve fitting
Fluorescence cross-correlation spectroscopy
business
Saturation (chemistry)
Algorithms
Excitation
Subjects
Details
- ISSN :
- 10833668
- Volume :
- 10
- Database :
- OpenAIRE
- Journal :
- Journal of Biomedical Optics
- Accession number :
- edsair.doi.dedup.....d0603b88fb01fc1302049d15e13d53f0