ArticleBiophysical journal1988
Polarized fluorescence photobleaching recovery for measuring rotational diffusion in solutions and membranes.
Article in Biophysical journal, 1988. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 87 citations in OpenAlex.
- Illuminating cellular architecture and dynamics with fluorescence polarization microscopy.Journal of cell science · 2024Review
- VAMP2 and synaptotagmin mobility in chromaffin granule membranes: implications for regulated exocytosis.Molecular biology of the cell · 2022Article
- Probing roto-translational diffusion of small anisotropic colloidal particles with a bright-field microscope.The European physical journal. E, Soft matter · 2021Article
- Visualization of expanding fusion pores in secretory cells.The Journal of general physiology · 2018Review
- Super-resolution dipole orientation mapping via polarization demodulation.Light, science & applications · 2016Article
- Photobleaching kinetics and time-integrated emission of fluorescent probes in cellular membranes.Molecules (Basel, Switzerland) · 2014Article
- Fluorescence lifetime excitation cytometry by kinetic dithering.Electrophoresis · 2014Article
- Protein mobility within secretory granules.Biophysical journal · 2014Article
- Rapid and quantitative imaging of excitation polarized fluorescence reveals ordered septin dynamics in live yeast.Biophysical journal · 2011Article
- Fluorescence anisotropy reveals order and disorder of protein domains in the nuclear pore complex.Biophysical journal · 2010Article
- Measuring rotational diffusion of MHC class I on live cells by polarized FPR.Biophysical chemistry · 2007Article
- Decreasing photobleaching by silver island films: application to muscle.Analytical biochemistry · 2007Article
- Intrasequence GFP in class I MHC molecules, a rigid probe for fluorescence anisotropy measurements of the membrane environment.Biophysical journal · 2003Article
- Lateral diffusion coefficients in membranes measured by resonance energy transfer and a new algorithm for diffusion in two dimensions.Biophysical journal · 2002Article
- Single-molecule anisotropy imaging.Biophysical journal · 1999Article
- Cell membrane orientation visualized by polarized total internal reflection fluorescence.Biophysical journal · 1999Article
- Chromatin dynamics in interphase nuclei and its implications for nuclear structure.The Journal of cell biology · 1997Article
- Photobleaching recovery and anisotropy decay of green fluorescent protein GFP-S65T in solution and cells: cytoplasmic viscosity probed by green fluorescent protein translational and rotational diffusion.Biophysical journal · 1997Article
- Cytoplasmic viscosity near the cell plasma membrane: translational diffusion of a small fluorescent solute measured by total internal reflection-fluorescence photobleaching recovery.Biophysical journal · 1996Article
- Myosin head rotation in muscle fibers measured using polarized fluorescence photobleaching recovery.Journal of fluorescence · 1995Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
A variation of fluorescence photobleaching recovery (FPR) suitable for measuring the rate of rotational molecular diffusion in solution and cell membranes is presented in theory and experimental practice for epi-illumination microscopy. In this technique, a brief flash of polarized laser light creates an anisotropic distribution of unbleached fluorophores which relaxes by rotational diffusion, leading to a time-dependent postbleach fluorescence. Polarized FPR (PFPR) is applicable to any time scales from seconds to microseconds. However, at fast (microsecond) time scales, a partial recovery independent of molecular orientation tends to obscure rotational effects. The theory here presents a method for overcoming this reversible photobleaching, and includes explicit results for practical geometries, fast wobble of fluorophores, and arbitrary bleaching depth. This variation of a polarized luminescence "pump-and-probe" technique is compared with prior ones and with "pump-only" time-resolved luminescence anisotropy decay methods. The technique is experimentally verified on small latex beads with a variety of diameters, common fluorophore labels, and solvent viscosities. Preliminary measurements on a protein (acetylcholine receptor) in the membrane of nondeoxygenated cells in live culture (rat myotubes) show a difference in rotational diffusion between clustered and nonclustered receptors. In most experiments, signal averaging, high laser power, and automated sample translation must be employed to achieve adequate statistical accuracy.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.