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

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Published on: November 23, 2021

Frapbot: An open-source application for FRAP data.

Robin Kohze¹, Cindy E J Dieteren², Werner J H Koopman¹

¹Department of Biochemistry (286), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, Nijmegen, GA 6525, The Netherlands.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|July 21, 2017

Summary

This summary is machine-generated.

Frapbot is a new, free open-source software for analyzing fluorescence recovery after photobleaching (FRAP) data. It offers both automated and manual analysis options for faster, more efficient biological research.

Keywords:

Frapbot R language fluorescence recovery after photobleaching pattern recognition web-application

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Area of Science:

Cell Biology
Biophysics
Biochemistry

Background:

Fluorescence Recovery After Photobleaching (FRAP) is a key technique for studying molecular dynamics in cells.
Manual analysis of FRAP data can be time-consuming and prone to user variability.
There is a need for accessible, automated tools to streamline FRAP data analysis.

Purpose of the Study:

To introduce Frapbot, a novel, free, open-source software application for FRAP data analysis.
To provide both automated and manual analysis capabilities for FRAP datasets.
To enhance the efficiency and reproducibility of FRAP experiments.

Main Methods:

Frapbot is developed in R, offering a web application interface.
It employs a pattern recognition algorithm for automated identification of relevant data columns and photobleaching time points.
The software visualizes raw data, residuals, normalization, and recovery half-times (t1/2) using user-definable fitting options.

Main Results:

Frapbot enables instant, batch-wise visualization of FRAP data and analysis results.
Automated analysis identifies relevant data and fitting parameters efficiently.
Fitted results are provided in .csv format within a .zip file for easy data handling.

Conclusions:

Frapbot offers a user-friendly, efficient solution for both automated and manual FRAP data analysis.
The software's open-source nature promotes accessibility and further development in the scientific community.
Frapbot facilitates robust analysis of molecular dynamics through FRAP experiments.