Determination of diffusion coefficients of fluorescently labelled molecules in lipid membranes using confocal scanning laser microscopy

N. Oncan, Michael Antony Player, Alistar Gordon MacDonald

Research output: Contribution to journalArticle

Abstract

A series of liposomes has been studied by the Fluorescence Recovery after Photobleaching technique using confocal scanning laser microscopy. Both mobile and immobile specimens of lipid membranes have been examined. The specimens consisted of liposomal suspensions incorporating mixtures of the phospholipids dioleoyl phosphatidylcholine (DOPC) and distearoyl phosphatidylcholine (DSPC) with the aminolipid dihexadecanol glycerophospho-ethanolamine (DHPE). The measurements reported provide information about the effective diffusion coefficients, which vary from 0.2 (.) 10(-10) cm(2)s(-1) to 3.2 (.) 10(-10) cm(2)s(-1). They were highly dependent on the ratio DSPC:DOPC. A simple theoretical model of 1-D diffusion was applied for the calculation of the effective diffusion coefficients. The measurements show that it is possible to use straightforward procedures with an unmodified commercial confocal microscope to determine diffusion coefficients of biological specimens.

Original languageEnglish
Pages (from-to)683-688
Number of pages5
JournalZeitschrift fur Naturforschung Section A-A Journal of Physical Sciences
Volume59
Issue number10
Publication statusPublished - 2004

Keywords

  • phospholipid
  • lateral diffusion
  • CSLM
  • DYNAMICS
  • MOBILITY
  • CELLS

Cite this

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title = "Determination of diffusion coefficients of fluorescently labelled molecules in lipid membranes using confocal scanning laser microscopy",
abstract = "A series of liposomes has been studied by the Fluorescence Recovery after Photobleaching technique using confocal scanning laser microscopy. Both mobile and immobile specimens of lipid membranes have been examined. The specimens consisted of liposomal suspensions incorporating mixtures of the phospholipids dioleoyl phosphatidylcholine (DOPC) and distearoyl phosphatidylcholine (DSPC) with the aminolipid dihexadecanol glycerophospho-ethanolamine (DHPE). The measurements reported provide information about the effective diffusion coefficients, which vary from 0.2 (.) 10(-10) cm(2)s(-1) to 3.2 (.) 10(-10) cm(2)s(-1). They were highly dependent on the ratio DSPC:DOPC. A simple theoretical model of 1-D diffusion was applied for the calculation of the effective diffusion coefficients. The measurements show that it is possible to use straightforward procedures with an unmodified commercial confocal microscope to determine diffusion coefficients of biological specimens.",
keywords = "phospholipid, lateral diffusion, CSLM, DYNAMICS, MOBILITY, CELLS",
author = "N. Oncan and Player, {Michael Antony} and MacDonald, {Alistar Gordon}",
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TY - JOUR

T1 - Determination of diffusion coefficients of fluorescently labelled molecules in lipid membranes using confocal scanning laser microscopy

AU - Oncan, N.

AU - Player, Michael Antony

AU - MacDonald, Alistar Gordon

PY - 2004

Y1 - 2004

N2 - A series of liposomes has been studied by the Fluorescence Recovery after Photobleaching technique using confocal scanning laser microscopy. Both mobile and immobile specimens of lipid membranes have been examined. The specimens consisted of liposomal suspensions incorporating mixtures of the phospholipids dioleoyl phosphatidylcholine (DOPC) and distearoyl phosphatidylcholine (DSPC) with the aminolipid dihexadecanol glycerophospho-ethanolamine (DHPE). The measurements reported provide information about the effective diffusion coefficients, which vary from 0.2 (.) 10(-10) cm(2)s(-1) to 3.2 (.) 10(-10) cm(2)s(-1). They were highly dependent on the ratio DSPC:DOPC. A simple theoretical model of 1-D diffusion was applied for the calculation of the effective diffusion coefficients. The measurements show that it is possible to use straightforward procedures with an unmodified commercial confocal microscope to determine diffusion coefficients of biological specimens.

AB - A series of liposomes has been studied by the Fluorescence Recovery after Photobleaching technique using confocal scanning laser microscopy. Both mobile and immobile specimens of lipid membranes have been examined. The specimens consisted of liposomal suspensions incorporating mixtures of the phospholipids dioleoyl phosphatidylcholine (DOPC) and distearoyl phosphatidylcholine (DSPC) with the aminolipid dihexadecanol glycerophospho-ethanolamine (DHPE). The measurements reported provide information about the effective diffusion coefficients, which vary from 0.2 (.) 10(-10) cm(2)s(-1) to 3.2 (.) 10(-10) cm(2)s(-1). They were highly dependent on the ratio DSPC:DOPC. A simple theoretical model of 1-D diffusion was applied for the calculation of the effective diffusion coefficients. The measurements show that it is possible to use straightforward procedures with an unmodified commercial confocal microscope to determine diffusion coefficients of biological specimens.

KW - phospholipid

KW - lateral diffusion

KW - CSLM

KW - DYNAMICS

KW - MOBILITY

KW - CELLS

M3 - Article

VL - 59

SP - 683

EP - 688

JO - Zeitschrift fur Naturforschung Section A-A Journal of Physical Sciences

JF - Zeitschrift fur Naturforschung Section A-A Journal of Physical Sciences

SN - 0932-0784

IS - 10

ER -