ArticleBiophysical journal2005
Fluorescence imaging of two-photon linear dichroism: cholesterol depletion disrupts molecular orientation in cell membranes.
Article in Biophysical journal, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 80 citations in OpenAlex.
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- Fluorescence lifetime imaging microscopy of flexible and rigid dyes probes the biophysical properties of synthetic and biological membranes.Biophysical journal · 2024Article
- Linear Dichroism Measurements for the Study of Protein-DNA Interactions.International journal of molecular sciences · 2023Review
- Unraveling the geometry of complex protein organizations by polarized fluorescence imaging.Biophysical journal · 2022Article
- Quantitative FRET Microscopy Reveals a Crucial Role of Cytoskeleton in Promoting PI(4,5)PInternational journal of molecular sciences · 2021Article
- Quantitative linear dichroism imaging of molecular processes in living cells made simple by open software tools.Communications biology · 2021Article
- High-dimensional super-resolution imaging reveals heterogeneity and dynamics of subcellular lipid membranes.Nature communications · 2020Article
- Advances of super-resolution fluorescence polarization microscopy and its applications in life sciences.Computational and structural biotechnology journal · 2020Review
- BODIPY-cholesterol can be reliably used to monitor cholesterol efflux from capacitating mammalian spermatozoa.Scientific reports · 2019Article
- Fundamental Limits on Measuring the Rotational Constraint of Single Molecules Using Fluorescence Microscopy.Physical review letters · 2019Article
- Fluorescence linear dichroism imaging for quantifying membrane order.Methods in molecular biology (Clifton, N.J.) · 2015Article
- Ultimate use of two-photon fluorescence microscopy to map orientational behavior of fluorophores.Biophysical journal · 2014Article
- Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure.Biophysical journal · 2013Article
- Mapping the local organization of cell membranes using excitation-polarization-resolved confocal fluorescence microscopy.Biophysical journal · 2013Article
- Probing the orientational distribution of dyes in membranes through multiphoton microscopy.Biophysical journal · 2012Article
- Probing orientational behavior of MHC class I protein and lipid probes in cell membranes by fluorescence polarization-resolved imaging.Biophysical journal · 2011Article
- Two-photon polarization microscopy reveals protein structure and function.Nature methods · 2011Article
- Peptide-induced domain formation in supported lipid bilayers: direct evidence by combined atomic force and polarized total internal reflection fluorescence microscopy.Biophysical journal · 2010Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The plasma membrane of cells is an ordered environment, giving rise to anisotropic orientation and restricted motion of molecules and proteins residing in the membrane. At the same time as being an organized matrix of defined structure, the cell membrane is heterogeneous and dynamic. Here we present a method where we use fluorescence imaging of linear dichroism to measure the orientation of molecules relative to the cell membrane. By detecting linear dichroism as well as fluorescence anisotropy, the orientation parameters are separated from dynamic properties such as rotational diffusion and homo energy transfer (energy migration). The sensitivity of the technique is enhanced by using two-photon excitation for higher photo-selection compared to single photon excitation. We show here that we can accurately image lipid organization in whole cell membranes and in delicate structures such as membrane nanotubes connecting two cells. The speed of our wide-field imaging system makes it possible to image changes in orientation and anisotropy occurring on a subsecond timescale. This is demonstrated by time-lapse studies showing that cholesterol depletion rapidly disrupts the orientation of a fluorophore located within the hydrophobic region of the cell membrane but not of a surface bound probe. This is consistent with cholesterol having an important role in stabilizing and ordering the lipid tails within the plasma membrane.
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Registered trials
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.