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Protein Dynamics in Living Cells

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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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Related Experiment Video

Updated: Jul 2, 2026

The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

Published on: October 12, 2010

Advanced application of CFSE for cellular tracking.

Jacek M Witkowski¹

¹Medical University of Gdańsk, Gdańsk, Poland.

Current Protocols in Cytometry

|September 5, 2008

Summary

This summary is machine-generated.

This study enhances cell tracking by quantifying lymphocyte proliferation dynamics. The improved method analyzes cell division, precursor numbers, and immune response efficiency without cell purification.

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The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

Published on: October 12, 2010

Isolation and Activation of Murine Lymphocytes

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Published on: October 30, 2016

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

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Published on: November 23, 2021

Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

The dividing-cell-tracking (DCT) technique uses CFSE staining to monitor lymphocyte division.
Existing DCT methods primarily focus on observing and counting cell generations.
A need exists for more detailed quantitative analysis of lymphocyte proliferation dynamics.

Purpose of the Study:

To extend the dividing-cell-tracking (DCT) cytometric technique.
To enable quantitative analysis of dynamic proliferation parameters in human lymphocytes.
To provide a method for assessing immune cell proliferative response efficiency.

Main Methods:

Utilizes supravital staining of lymphocytes with CFSE.
Involves tracking cell division and generations over time.
Applies calculations to cytometrically discernible subpopulations without prior purification.

Main Results:

Determines the duration of the pre-division transition period (G0 to G1).
Quantifies the time of a single cell division.
Calculates the average number of divisions per cell and the number of effective precursors.
Enables analysis of various lymphocyte sources and subpopulations.

Conclusions:

The enhanced DCT method provides dynamic proliferation parameters for in vitro human lymphocytes.
It offers a powerful tool for detailed analysis of immune cell proliferative responses.
The technique's ability to analyze unpurified populations increases its utility in immunological studies.