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Updated: Jun 21, 2025
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How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
Published on: January 7, 2019
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Photoactivatable Blue Fluorescent Protein.
Paul Gaytán1, Abigail Roldán-Salgado1
1Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos 62210, Mexico.
ACS Omega
|July 8, 2024
Summary
Researchers developed a novel photoactivatable blue fluorescent protein (PA-BFP) from SumireF. This new tool enables advanced cellular imaging and tracking of biological processes with existing microscopy technology.
Area of Science:
- Biochemistry
- Molecular Biology
- Microscopy
Background:
- Photoactivatable and photoswitchable fluorescent proteins (FPs) are crucial for super-resolution microscopy.
- Existing FPs have limitations in specific applications and require advanced imaging setups.
Purpose of the Study:
- To create a novel photoactivatable blue fluorescent protein (PA-BFP).
- To establish a new genetically encoded reporter for cellular imaging compatible with existing technologies.
Main Methods:
- A Y66H mutation was introduced into the chromophore of the violet fluorescent protein SumireF.
- The resulting PA-BFP was characterized for its activation properties and spectral characteristics.
Main Results:
- The first photoactivatable blue fluorescent protein (PA-BFP) was successfully created.
- PA-BFP is activated by UV transilluminators (302/365 nm) and sunlight.
- Its spectral properties (excitation/emission ~358/445 nm) match DAPI, a common blue stain.
Conclusions:
- PA-BFP is a versatile tool for cellular biology, leveraging established microscopy techniques.
- It can be used with other FPs for multicolor labeling and simultaneous tracking of multiple proteins.
- This advance facilitates elucidation of complex biological processes using advanced imaging.
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