Quantitative imaging of molecular order in lipid membranes using two-photon fluorescence polarimetry

Alicja Gasecka; Tsai Jung Han; Cyril Favard; Bong Rae Cho; Sophie Brasselet

doi:10.1016/j.bpj.2009.08.052

Quantitative imaging of molecular order in lipid membranes using two-photon fluorescence polarimetry

Alicja Gasecka, Tsai Jung Han, Cyril Favard, Bong Rae Cho, Sophie Brasselet

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59 Citations (Scopus)

Abstract

We present a polarimetric two-photon microscopy technique to quantitatively image the local static molecular orientational behavior in lipid and cell membranes. This approach, based on a tunable excitation polarization state complemented by a polarized readout, is easily implementable and does not require hypotheses on the molecular angular distribution such as its mean orientation, which is a main limitation in traditional fluorescence anisotropy measurements. The method is applied to the investigation of the molecular angular distribution in giant unilamellar vesicles formed by liquid-ordered and liquid-disordered micro-domains, and in COS-7 cell membranes. The highest order contrast between ordered and disordered domains is obtained for dyes locating within the membrane acyl chains.

Original language	English
Pages (from-to)	2854-2862
Number of pages	9
Journal	Biophysical Journal
Volume	97
Issue number	10
DOIs	https://doi.org/10.1016/j.bpj.2009.08.052
Publication status	Published - 2009 Nov 15

Bibliographical note

Funding Information:
This work was supported by the French Agence Nationale de la Recherche under program No. JC 2007 (project NLO-Shaping grant No. JC07-195504), the European Commission through the Human Potential Program Marie-Curie RTN NANOMATCH (grant No. MRTN-CT-2006-035884), and the Korea Science and Engineering Foundation grant (No. R0A-2007-000-20027-0) funded by the Korea Ministry of Education, Science, and Technology.

ASJC Scopus subject areas

Biophysics

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10.1016/j.bpj.2009.08.052

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title = "Quantitative imaging of molecular order in lipid membranes using two-photon fluorescence polarimetry",

abstract = "We present a polarimetric two-photon microscopy technique to quantitatively image the local static molecular orientational behavior in lipid and cell membranes. This approach, based on a tunable excitation polarization state complemented by a polarized readout, is easily implementable and does not require hypotheses on the molecular angular distribution such as its mean orientation, which is a main limitation in traditional fluorescence anisotropy measurements. The method is applied to the investigation of the molecular angular distribution in giant unilamellar vesicles formed by liquid-ordered and liquid-disordered micro-domains, and in COS-7 cell membranes. The highest order contrast between ordered and disordered domains is obtained for dyes locating within the membrane acyl chains.",

author = "Alicja Gasecka and Han, {Tsai Jung} and Cyril Favard and Cho, {Bong Rae} and Sophie Brasselet",

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AU - Gasecka, Alicja

AU - Han, Tsai Jung

AU - Favard, Cyril

AU - Cho, Bong Rae

AU - Brasselet, Sophie

N1 - Funding Information: This work was supported by the French Agence Nationale de la Recherche under program No. JC 2007 (project NLO-Shaping grant No. JC07-195504), the European Commission through the Human Potential Program Marie-Curie RTN NANOMATCH (grant No. MRTN-CT-2006-035884), and the Korea Science and Engineering Foundation grant (No. R0A-2007-000-20027-0) funded by the Korea Ministry of Education, Science, and Technology.

PY - 2009/11/15

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N2 - We present a polarimetric two-photon microscopy technique to quantitatively image the local static molecular orientational behavior in lipid and cell membranes. This approach, based on a tunable excitation polarization state complemented by a polarized readout, is easily implementable and does not require hypotheses on the molecular angular distribution such as its mean orientation, which is a main limitation in traditional fluorescence anisotropy measurements. The method is applied to the investigation of the molecular angular distribution in giant unilamellar vesicles formed by liquid-ordered and liquid-disordered micro-domains, and in COS-7 cell membranes. The highest order contrast between ordered and disordered domains is obtained for dyes locating within the membrane acyl chains.

AB - We present a polarimetric two-photon microscopy technique to quantitatively image the local static molecular orientational behavior in lipid and cell membranes. This approach, based on a tunable excitation polarization state complemented by a polarized readout, is easily implementable and does not require hypotheses on the molecular angular distribution such as its mean orientation, which is a main limitation in traditional fluorescence anisotropy measurements. The method is applied to the investigation of the molecular angular distribution in giant unilamellar vesicles formed by liquid-ordered and liquid-disordered micro-domains, and in COS-7 cell membranes. The highest order contrast between ordered and disordered domains is obtained for dyes locating within the membrane acyl chains.

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Quantitative imaging of molecular order in lipid membranes using two-photon fluorescence polarimetry

Abstract

Bibliographical note

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