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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

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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Related Experiment Video

Updated: Mar 15, 2026

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Published on: December 17, 2015

Phase correlation imaging of unlabeled cell dynamics.

Lihong Ma^1,2, Gannavarpu Rajshekhar¹, Ru Wang¹

¹Quantitative Light Imaging Laboratory, Department of Electrical and Computer Engineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.

Scientific Reports

|September 13, 2016

Summary

Phase Correlation Imaging (PCI) offers a label-free method to study cell dynamics by analyzing intracellular mass transport. This technique quantitatively maps diffusion coefficients and assesses actin polymerization, distinguishing cell states like senescence.

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Area of Science:

Cell Biology
Biophysics
Quantitative Imaging

Background:

Studying cell dynamics often requires exogenous markers, which can introduce limitations.
Understanding intracellular mass transport is crucial for cell function.
Distinguishing between different cell states, such as senescent and quiescent, is challenging.

Purpose of the Study:

To introduce Phase Correlation Imaging (PCI) as a novel, label-free technique for studying cell dynamics.
To demonstrate PCI's capability in quantitatively assessing intracellular mass transport and diffusion.
To explore PCI's application in evaluating actin polymerization dynamics and cell state differentiation.

Main Methods:

Utilized quantitative phase imaging time-lapse data.
Employed phase correlation analysis to generate correlation time maps.
Applied PCI to live cells and standard Brownian particles for validation.

Main Results:

PCI successfully generated quantitative diffusion coefficient maps for live cells and Brownian particles.
The technique revealed that cyto-D treatment, which blocks actin polymerization, primarily affects dynamics at large spatial scales.
PCI demonstrated the ability to differentiate between senescent and quiescent cells without specific markers.

Conclusions:

Phase Correlation Imaging is a powerful, label-free tool for spatially-resolved analysis of cell dynamics.
PCI provides quantitative insights into intracellular mass transport, diffusion, and actin polymerization.
PCI is anticipated to complement existing fluorescence-based methods for advanced cell function studies.