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Related Experiment Video

Updated: Jun 5, 2026

Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer
07:54

Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer

Published on: October 15, 2015

A nonfitting method using a spatial sine window transform for inhomogeneous effective-diffusion measurements by FRAP.

Darya Y Orlova1, Eva Bártová, Valeri P Maltsev

  • 1Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic.

Biophysical Journal
|January 20, 2011
PubMed
Summary

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This study presents a novel nonfitting method to determine effective diffusion coefficients in complex cellular environments. The technique analyzes fluorescence recovery patterns, simplifying the study of protein mobility in inhomogeneous regions like cell nuclei.

Area of Science:

  • Cellular Biophysics
  • Molecular Biology
  • Biochemistry

Background:

  • Accurately measuring protein diffusion in inhomogeneous cellular environments, especially with ligand-receptor interactions, is analytically challenging.
  • Conventional methods for analyzing fluorescence recovery after photobleaching (FRAP) data often require complex mathematical solutions for diffusion-reaction equations.

Purpose of the Study:

  • To introduce a nonfitting mathematical method for evaluating averaged effective diffusion coefficients.
  • To enable the analysis of protein mobility within complex, inhomogeneous cellular regions like the nucleus.

Main Methods:

  • Utilized a nonfitting mathematical processing approach on the entire cellular 2D spatial pattern of recovered fluorescence.
  • Employed spatially and temporally resolved measurements via fluorescence recovery after photobleaching (FRAP).

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From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope
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Last Updated: Jun 5, 2026

Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer
07:54

Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer

Published on: October 15, 2015

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells
05:56

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

Published on: November 12, 2020

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope
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From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

Published on: October 9, 2014

  • Collected 2D fluorescence images using laser-scanning confocal microscopy.

    • Main Results:

    • Successfully demonstrated a nonfitting method to calculate averaged effective diffusion coefficients.
    • Applied the method to estimate the mobility of green fluorescent protein-tagged heterochromatin protein 1 (HP1) in mouse embryonic fibroblast nuclei.
    • The method bypasses the need for solving complex diffusion-reaction equations.

    Conclusions:

    • The developed nonfitting method offers a more tractable approach to determining effective diffusion constants in complex biological systems.
    • This technique is particularly advantageous for studying protein dynamics in highly inhomogeneous cellular compartments, such as the nucleus with its protein foci and chromatin domains.