ArticleBiomedical optics express2012
Focal switching of photochromic fluorescent proteins enables multiphoton microscopy with superior image contrast.
Article in Biomedical optics express, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging.Nature biotechnology · 2022Article
- Two-photon probes for in vivo multicolor microscopy of the structure and signals of brain cells.Brain structure & function · 2018Review
- Two-photon-like microscopy with orders-of-magnitude lower illumination intensity via two-step fluorescence.Nature communications · 2015Article
- A fast- and positively photoswitchable fluorescent protein for ultralow-laser-power RESOLFT nanoscopy.Nature methods · 2015Article
- Reversibly switchable fluorescence microscopy with enhanced resolution and image contrast.Journal of biomedical optics · 2014Article
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Authors and funding
4 authors.
Funding
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Abstract
Probing biological structures and functions deep inside live organisms with light is highly desirable. Among the current optical imaging modalities, multiphoton fluorescence microscopy exhibits the best contrast for imaging scattering samples by employing a spatially confined nonlinear excitation. However, as the incident laser power drops exponentially with imaging depth into the sample due to the scattering loss, the out-of-focus background eventually overwhelms the in-focus signal, which defines a fundamental imaging-depth limit. Herein we significantly improve the image contrast for deep scattering samples by harnessing reversibly switchable fluorescent proteins (RSFPs) which can be cycled between bright and dark states upon light illumination. Two distinct techniques, multiphoton deactivation and imaging (MPDI) and multiphoton activation and imaging (MPAI), are demonstrated on tissue phantoms labeled with Dronpa protein. Such a focal switch approach can generate pseudo background-free images. Conceptually different from wave-based approaches that try to reduce light scattering in turbid samples, our work represents a molecule-based strategy that focused on imaging probes.
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