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Updated: Jun 4, 2026
12:20
Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
Imaging Real-Time Gene Expression in Living Systems with Single-Transcript Resolution: Single mRNA Particle Tracking
Amber L Wells1, John S Condeelis, Robert H Singer
1Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
CSH Protocols
|March 2, 2011
Summary
This protocol details using ImageJ software to analyze fluorescent mRNA particle dynamics within cells from time-lapse microscopy. The method generates maximum intensity projections and kymographs for visualization.
Area of Science:
- Cell biology
- Biophysics
- Microscopy image analysis
Background:
- Understanding intracellular transport is crucial for cell function.
- Analyzing the movement of messenger RNA (mRNA) particles provides insights into gene expression regulation.
- Quantitative analysis of particle dynamics often requires specialized software tools.
Purpose of the Study:
- To provide a protocol for analyzing fluorescent mRNA particle dynamics in cells.
- To demonstrate the utility of ImageJ software for quantitative microscopy.
- To generate visualizations like maximum intensity projections and kymographs.
Main Methods:
- Utilizing ImageJ, an open-source image processing program.
- Analyzing time-lapse movie frames or image stacks of fluorescent mRNA particles.
- Generating maximum intensity projections to visualize particle distribution.
- Creating kymographs to analyze particle movement over time.
Main Results:
- The protocol enables visualization of fluorescent mRNA particle movement within cellular environments.
- Maximum intensity projections highlight the spatial distribution of particles.
- Kymographs allow for the assessment of particle velocity and directionality.
Conclusions:
- ImageJ is a powerful and accessible tool for analyzing particle dynamics in live-cell imaging.
- The described methods facilitate quantitative studies of mRNA transport and localization.
- This protocol can be adapted for analyzing other types of intracellular particles.
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