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Related Experiment Video

Updated: Jul 2, 2025

Photoconversion of Purified Fluorescent Proteins and Dual-probe Optical Highlighting in Live Cells

Photoconversion of Purified Fluorescent Proteins and Dual-probe Optical Highlighting in Live Cells

Published on: June 26, 2010

Using ER-Targeted Photoconvertible Fluorescent Proteins in Living Plant Cells.

Jaideep Mathur¹, Puja Puspa Ghosh²

¹Laboratory of Plant Development & Interactions, Department of Molecular & Cellular Biology, University of Guelph, Guelph, ON, Canada. jmathur@uoguelph.ca.

Methods in Molecular Biology (Clifton, N.J.)

|February 27, 2024

Summary

This study details live-cell imaging using photoconvertible fluorescent proteins (pcFPs) targeted to the endoplasmic reticulum (ER). We provide methods and troubleshooting for studying ER dynamics and organelle interactions.

Keywords:

Dendra Endoplasmic reticulum Kaede Live-cell imaging Organellar dynamics Photoconvertible fluorescent proteins mEos mMaple

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Area of Science:

Cell Biology
Biophysics
Microscopy

Background:

Photoconvertible fluorescent proteins (pcFPs) allow for multicolor labeling within single cells.
Endoplasmic reticulum (ER) targeted pcFP probes are valuable for studying ER dynamics.
Understanding ER interactions with other organelles is crucial in cell biology.

Purpose of the Study:

To describe live-cell imaging procedures using ER-targeted pcFP probes.
To provide solutions for potential experimental problems.
To suggest improvements for ER live-cell imaging experiments.

Main Methods:

Utilizing ER-targeted pcFP-based probes for live-cell imaging.
Implementing photoconversion techniques for differential labeling.
Performing live-cell microscopy to observe ER dynamics.

Main Results:

Successful application of ER-targeted pcFP probes in live-cell imaging.
Identification of common experimental challenges and their resolutions.
Demonstration of optimized protocols for enhanced imaging quality.

Conclusions:

ER-targeted pcFP probes are effective tools for live-cell imaging of the endoplasmic reticulum.
The described methods and troubleshooting guide facilitate robust ER dynamics studies.
This work enhances the utility of pcFPs for investigating organelle interactions.