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Related Concept Videos

Protein Diffusion in the Membrane01:24

Protein Diffusion in the Membrane

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Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...
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Facilitated Diffusion01:16

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The plasma membrane, a critical structure in cellular biology, houses an array of transporters, or carrier proteins, interspersed within its lipid bilayer. These proteins play a crucial role in solute transport through facilitated diffusion, a form of passive diffusion that uses transporters to move the molecules across the membrane.
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Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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In column chromatography, when an analyte is introduced as a narrow band at the top of the column, the solutes begin to separate and broaden, developing a Gaussian profile. This broadening occurs due to various factors, such as longitudinal diffusion.
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The chemical and physical properties of plasma membranes cause them to be selectively permeable. Since plasma membranes have both hydrophobic and hydrophilic regions, substances need to be able to transverse both regions. The hydrophobic area of membranes repels substances such as charged ions. Therefore, such substances need special membrane proteins to cross a membrane successfully. In  facilitated transport, also known as facilitated diffusion, molecules and ions travel across a...
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Facilitated Transport01:19

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The chemical and physical properties of plasma membranes cause them to be selectively permeable. Since plasma membranes have both hydrophobic and hydrophilic regions, substances need to be able to transverse both regions. The hydrophobic area of membranes repels substances such as charged ions. Therefore, such substances need special membrane proteins to cross a membrane successfully. In  facilitated transport, also known as facilitated diffusion, molecules and ions travel across a...
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Related Experiment Video

Updated: Dec 9, 2025

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells
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Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells

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Variations in Plasma Membrane Topography Can Explain Heterogenous Diffusion Coefficients Obtained by Fluorescence

Astrid Gesper1, Stefan Wennmalm2, Philipp Hagemann1

  • 1RUBION, Ruhr-Universität Bochum, Bochum, Germany.

Frontiers in Cell and Developmental Biology
|September 9, 2020
PubMed
Summary

Cell membrane topography significantly impacts fluorescence correlation spectroscopy (FCS) diffusion measurements. Smoother membrane areas, like above the nucleus, show faster diffusion, highlighting the need to consider surface variations.

Keywords:
diffusionfluorescence correlation spectroscopymembrane topographyplasma membranescanning ion conductance microscopy

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Area of Science:

  • Cell biology
  • Biophysics
  • Membrane dynamics

Background:

  • Fluorescence correlation spectroscopy (FCS) is widely used to measure diffusion in cell membranes, particularly the plasma membrane.
  • Observed diffusion coefficients in plasma membranes often show considerable variability within the same cell type or even within a single cell.

Purpose of the Study:

  • To investigate if variations in cell membrane topography explain the spread in FCS diffusion measurements.
  • To determine the influence of membrane surface roughness on diffusion dynamics.

Main Methods:

  • Utilized fluorescence correlation spectroscopy (FCS) to measure the diffusion of the membrane dye DiI.
  • Employed live-cell scanning ion conductance microscopy (SICM) to map cell surface topography.
  • Performed FCS modeling with simulated diffusion on SICM-derived surfaces.

Main Results:

  • Diffusion of DiI was faster in the apical plasma membrane above the nucleus compared to above the cytoplasm.
  • SICM revealed that the plasma membrane above the nucleus is smoother than other areas, correlating with faster diffusion.
  • FCS modeling confirmed that topography variations can lead to apparent anomalous diffusion.
  • Variations in the membrane marker DiD within the FCS focal volume correlated with diffusion time differences, unlike transmembrane or lipid-anchored proteins.

Conclusions:

  • Cell surface topography is a critical factor influencing FCS measurements of membrane diffusion.
  • The uneven distribution of membrane roughness must be considered when interpreting FCS data from cellular membranes.
  • Topographical variations can account for apparent anomalies in diffusion observed via FCS.