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Fluorescence imagin...
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Benninger, R. K. P.
(author)
Fluorescence imaging of two-photon linear dichroism : Cholesterol depletion disrupts molecular orientation in cell membranes
- Article/chapterEnglish2005
Publisher, publication year, extent ...
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Elsevier BV,2005
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printrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:kth-14444
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https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-14444URI
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https://doi.org/10.1529/biophysj.104.050096DOI
Supplementary language notes
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Language:English
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Summary in:English
Part of subdatabase
Classification
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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QC 20100525
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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.
Subject headings and genre
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dipyrrometheneboron difluoride bodipy
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multifocal multiphoton microscopy
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lipid rafts
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plasma-membrane
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bilayer-membranes
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mammalian-cells
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chain-length
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living cells
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immune cells
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nk cells
Added entries (persons, corporate bodies, meetings, titles ...)
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Önfelt, Björn
(author)
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Neil, M. A. A.
(author)
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Davis, D. M.
(author)
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French, P. M. W.
(author)
Related titles
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In:Biophysical Journal: Elsevier BV88:1, s. 609-6220006-34951542-0086
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