Enhanced-contrast two-photon optogenetic pH sensing and pH-resolved brain imaging

A. Chebotarev; M. Pochechuev; A. Lanin; I. Kelmanson; D. Kotova; E. Fetisova; A. Panova; D. Bilan; A. Fedotov; V. Belousov; A. Zheltikov

Статья

Enhanced-contrast two-photon optogenetic pH sensing and pH-resolved brain imaging

A. Chebotarev, M. Pochechuev, A. Lanin, I. Kelmanson, D. Kotova, E. Fetisova, A. Panova, D. Bilan, A. Fedotov, V. Belousov, A. Zheltikov,

2021

https://doi.org/10.1002/jbio.202000301

We present experiments on cell cultures and brain slices that demonstrate two-photon optogenetic pH sensing and pH-resolved brain imaging using a laser driver whose spectrum is carefully tailored to provide the maximum contrast of a ratiometric two-photon fluorescence readout from a high-brightness genetically encoded yellow-fluorescent-protein-based sensor, SypHer3s. Two spectrally isolated components of this laser field are set to induce two-photon-excited fluorescence (2PEF) by driving SypHer3s through one of two excitation pathways—via either the protonated or deprotonated states of its chromophore. With the spectrum of the laser field accurately adjusted for a maximum contrast of these two 2PEF signals, the ratio of their intensities is shown to provide a remarkably broad dynamic range for pH measurements, enabling high-contrast optogenetic deep-brain pH sensing and pH-resolved 2PEF imaging within a vast class of biological systems, ranging from cell cultures to the living brain.

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1. Version of Record от 2021-03-01

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Об авторах

A. Chebotarev
Lomonosov Moscow State University
M. Pochechuev
Lomonosov Moscow State University
A. Lanin
Lomonosov Moscow State University, National University of Science & Technology (MISIS), Kazan National Research Technical University named after A. N. Tupolev -KAI
I. Kelmanson
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Pirogov Russian National Research Medical University (RNRMU)
D. Kotova
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
E. Fetisova
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
A. Panova
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
D. Bilan
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Pirogov Russian National Research Medical University (RNRMU)
A. Fedotov
Lomonosov Moscow State University, National University of Science & Technology (MISIS), Kazan National Research Technical University named after A. N. Tupolev -KAI, National University of Science & Technology (MISIS)
V. Belousov
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Pirogov Russian National Research Medical University (RNRMU), Federal Center of Brain Research and Neurotechnologies of the Federal Medical Biological Agency
A. Zheltikov
Lomonosov Moscow State University, National University of Science & Technology (MISIS), Kazan National Research Technical University named after A. N. Tupolev -KAI, Texas A&M University

Название журнала

Journal of Biophotonics

Том

Выпуск

Финансирующая организация

Welch Foundation

Номер гранта

A‐1801‐20180324

Тип документа

journal article

Тип лицензии Creative Commons

CC BY

Правовой статус документа

Свободная лицензия

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scopus

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Статья

Enhanced-contrast two-photon optogenetic pH sensing and pH-resolved brain imaging

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