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Biochimica et Biophysica Acta (BBA) - Biomembranes
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Biochimica et Biophysica Acta (BBA) - Biomembranes
Article . 2010
License: Elsevier Non-Commercial
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Biochimica et Biophysica Acta (BBA) - Biomembranes
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Multiphoton excitation fluorescence microscopy in planar membrane systems

Authors: Brewer, Jonathan R.; id_orcid 0000-0002-3444-1715; Bernardino de la Serna, Jorge; Wagner, Kerstin; Bagatolli, Luis;

Multiphoton excitation fluorescence microscopy in planar membrane systems

Abstract

The feasibility of applying multiphoton excitation fluorescence microscopy-related techniques in planar membrane systems, such as lipid monolayers at the air-water interface (named Langmuir films), is presented and discussed in this paper. The non-linear fluorescence microscopy approach, allows obtaining spatially and temporally resolved information by exploiting the fluorescent properties of particular fluorescence probes. For instance, the use of environmental sensitive probes, such as LAURDAN, allows performing measurements using the LAURDAN generalized polarization function that in turn is sensitive to the local lipid packing in the membrane. The fact that LAURDAN exhibit homogeneous distribution in monolayers, particularly in systems displaying domain coexistence, overcomes a general problem observed when "classical" fluorescence probes are used to label Langmuir films, i.e. the inability to obtain simultaneous information from the two coexisting membrane regions. Also, the well described photoselection effect caused by excitation light on LAURDAN allows: (i) to qualitative infer tilting information of the monolayer when liquid condensed phases are present and (ii) to provide high contrast to visualize 3D membranous structures at the film's collapse pressure. In the last case, computation of the LAURDAN GP function provides information about lipid packing in these 3D structures. Additionally, LAURDAN GP values upon compression in monolayers were compared with those obtained in compositionally similar planar bilayer systems. At similar GP values we found, for both DOPC and DPPC, a correspondence between the molecular areas reported in monolayers and bilayers. This correspondence occurs when the lateral pressure of the monolayer is 26+/-2 mN/m and 28+/-3 mN/m for DOPC and DPPC, respectively.

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Denmark
Related Organizations
Keywords

1,2-Dipalmitoylphosphatidylcholine, Biophysics, Langmuir films, Membranes, Artificial, Cell Biology, Biochemistry, Phase Transition, Planar supported membranes, Microscopy, Fluorescence, Multiphoton, 2-Naphthylamine, Phosphatidylcholines, LAURDAN, Two-photon excitation microscopy, Laurates

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
59
Top 10%
Top 10%
Top 10%
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