ArticleBiomedical optics express2022
Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors.
Article in Biomedical optics express, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Methods, Trade-offs, and Opportunities in High-speed Optical Microscopy for Neural Voltage imaging.Nature photonics · 2026Article
- Natural Product-Based Upconversion-Downshifting Photosensitizers in Photodynamic Therapy.Pharmaceuticals (Basel, Switzerland) · 2026Review
- Optical constraints on two-photon voltage imaging.Neurophotonics · 2024Article
- Optical constraints on two-photon voltage imaging.bioRxiv : the preprint server for biology · 2024Article
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Authors and funding
7 authors.
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Abstract
Fluorescent proteins are excited by light that is polarized parallel to the dipole axis of the chromophore. In two-photon microscopy, polarized light is used for excitation. Here we reveal surprisingly strong polarization sensitivity in a class of genetically encoded, GPCR-based neurotransmitter sensors. In tubular structures such as dendrites, this effect led to a complete loss of membrane signal in dendrites running parallel to the polarization direction of the excitation beam. To reduce the sensitivity to dendritic orientation, we designed an optical device that generates interleaved pulse trains of orthogonal polarization. The passive device, which we inserted in the beam path of an existing two-photon microscope, removed the strong direction bias from fluorescence and second-harmonic (SHG) images. We conclude that for optical measurements of transmitter concentration with GPCR-based sensors, orthogonally polarized excitation is essential.
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