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Varied Length Stokes Shift BODIPY-Based Fluorophores for Multicolor Microscopy
Amy M Bittel1, Ashley M Davis1, Lei Wang1
1Biomedical Engineering Department, Oregon Health & Science University, Portland, OR, 97201, USA.
Scientific Reports
|March 17, 2018
Summary
Researchers developed novel boron-dipyrromethene (BODIPY)-based fluorophores to overcome limitations in multicolor microscopy. These new probes enable advanced imaging, allowing for more colors to be visualized in a single sample.
Area of Science:
- Chemical Biology
- Microscopy
- Spectroscopy
Background:
- Current fluorophore technology limits multicolor microscopy due to short Stokes shifts, restricting detection to 4-5 colors.
- Commercial fluorophores are optimized for conventional microscopy, not ideal for highly multiplexed applications.
Purpose of the Study:
- To synthesize and screen novel boron-dipyrromethene (BODIPY)-based fluorophores for multicolor microscopy.
- To develop probes with varied Stokes shifts for enhanced multiplexing capabilities.
Main Methods:
- Synthesis of a library of 95 novel BODIPY-based fluorophores.
- Screening of photophysical, optical, and spectral properties.
- Confocal laser scanning microscopy with single excitation.
Main Results:
- A subset of BODIPY fluorophores with varied Stokes shifts was identified.
- Demonstrated the first five-color image using single excitation with these novel fluorophores.
- Showcased potential for up to 9-10 colors with linear unmixing.
Conclusions:
- Novel BODIPY fluorophores significantly advance multicolor microscopy capabilities.
- These probes overcome limitations of existing fluorophores for highly multiplexed imaging.
- Future applications include enhanced subcellular system visualization.
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