Related Experiment Video
Updated: May 25, 2026
11:11
Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
Published on: August 15, 2013
Flow cytometry of fluorescent proteins.
William G Telford1, Teresa Hawley, Fedor Subach
1Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. telfordw@mail.nih.gov
Methods (San Diego, Calif.)
|February 2, 2012
Summary
Fluorescent proteins are essential for biomedical research. This review details their use in flow cytometry, focusing on excitation, emission, and equipment needs for optimal results.
Area of Science:
- Biomedical Research
- Cellular Biology
- Biotechnology
Background:
- Fluorescent proteins (FPs) are indispensable tools in modern biomedical research.
- Their application spans diverse fields, including molecular and cellular biology.
- Advances in FP technology have significantly enhanced imaging and detection capabilities.
Purpose of the Study:
- To provide a comprehensive review of techniques for utilizing fluorescent proteins in flow cytometry.
- To detail the specific excitation and emission requirements for various fluorescent proteins.
- To outline the essential equipment necessary for optimal fluorescent protein-based flow cytometry.
Main Methods:
- Review of current literature and established protocols for fluorescent protein application in flow cytometry.
- Analysis of spectral properties (excitation and emission) of commonly used fluorescent proteins.
- Identification of critical instrumentation and setup parameters for effective flow cytometry.
Main Results:
- Detailed compilation of excitation and emission spectra for key fluorescent proteins.
- Guidelines for selecting appropriate fluorescent proteins based on experimental design and equipment capabilities.
- Recommendations for optimizing flow cytometer settings for enhanced signal detection and reduced spectral overlap.
Conclusions:
- Effective use of fluorescent proteins in flow cytometry requires careful consideration of spectral properties and equipment.
- Proper selection and implementation of fluorescent proteins can significantly improve data quality and experimental outcomes.
- This review serves as a practical guide for researchers aiming to leverage fluorescent proteins in flow cytometry applications.
Related Concept Videos
01:23Flow Cytometry
The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...
In...
01:19Protein 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...
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...