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Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors

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Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors. / Pulin, Mauro; Stockhausen, Kilian E.; Masseck, Olivia A.; Kubitschke, Martin; Busse, Björn; Wiegert, J. Simon; Oertner, Thomas G.

In: BIOMED OPT EXPRESS, Vol. 13, No. 2, 01.02.2022, p. 777-790.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Pulin, M, Stockhausen, KE, Masseck, OA, Kubitschke, M, Busse, B, Wiegert, JS & Oertner, TG 2022, 'Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors', BIOMED OPT EXPRESS, vol. 13, no. 2, pp. 777-790. https://doi.org/10.1364/BOE.448760

APA

Pulin, M., Stockhausen, K. E., Masseck, O. A., Kubitschke, M., Busse, B., Wiegert, J. S., & Oertner, T. G. (2022). Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors. BIOMED OPT EXPRESS, 13(2), 777-790. https://doi.org/10.1364/BOE.448760

Vancouver

Pulin M, Stockhausen KE, Masseck OA, Kubitschke M, Busse B, Wiegert JS et al. Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors. BIOMED OPT EXPRESS. 2022 Feb 1;13(2):777-790. https://doi.org/10.1364/BOE.448760

Bibtex

@article{ab386095e32e4ae79c2097ed280b4b75,
title = "Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors",
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.",
author = "Mauro Pulin and Stockhausen, {Kilian E.} and Masseck, {Olivia A.} and Martin Kubitschke and Bj{\"o}rn Busse and Wiegert, {J. Simon} and Oertner, {Thomas G.}",
note = "Publisher Copyright: {\textcopyright} 2022 OSA - The Optical Society. All rights reserved.",
year = "2022",
month = feb,
day = "1",
doi = "10.1364/BOE.448760",
language = "English",
volume = "13",
pages = "777--790",
journal = "BIOMED OPT EXPRESS",
issn = "2156-7085",
publisher = "The Optical Society",
number = "2",

}

RIS

TY - JOUR

T1 - Orthogonally-polarized excitation for improved two-photon and second-harmonic-generation microscopy, applied to neurotransmitter imaging with GPCR-based sensors

AU - Pulin, Mauro

AU - Stockhausen, Kilian E.

AU - Masseck, Olivia A.

AU - Kubitschke, Martin

AU - Busse, Björn

AU - Wiegert, J. Simon

AU - Oertner, Thomas G.

N1 - Publisher Copyright: © 2022 OSA - The Optical Society. All rights reserved.

PY - 2022/2/1

Y1 - 2022/2/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85123443939&partnerID=8YFLogxK

U2 - 10.1364/BOE.448760

DO - 10.1364/BOE.448760

M3 - SCORING: Journal article

AN - SCOPUS:85123443939

VL - 13

SP - 777

EP - 790

JO - BIOMED OPT EXPRESS

JF - BIOMED OPT EXPRESS

SN - 2156-7085

IS - 2

ER -