Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential

Okhil K. Nag; Ji Eun Jeong; Van Sang Le; Eunkeu Oh; Han Young Woo; James B. Delehanty

doi:10.1111/php.13233

Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential

Okhil K. Nag, Ji Eun Jeong, Van Sang Le, Eunkeu Oh, Han Young Woo, James B. Delehanty

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

We report a Förster resonance energy transfer (FRET)-based imaging ensemble for the visualization of membrane potential in living cells. A water-soluble poly(fluorene-cophenylene) conjugated polyelectrolyte (FsPFc10) serves as a FRET donor to a voltage-sensitive dye acceptor (FluoVolt^™). We observe FRET between FsPFc10 and FluoVolt^™, where the enhancement in FRET-sensitized emission from FluoVolt^™ is measured at various donor/acceptor ratios. At a donor/acceptor ratio of 1, the excitation of FluoVolt^™ in a FRET configuration results in a three-fold enhancement in its fluorescence emission (compared to when it is excited directly). FsPFc10 efficiently labels the plasma membrane of HEK 293T/17 cells and remains resident with minimal cellular internalization for ~ 1.5 h. The successful plasma membrane-associated colabeling of the cells with the FsPFc10-FluoVolt^™ donor-acceptor pair is confirmed by dual-channel confocal imaging. Importantly, cells labeled with FsPFc10 show excellent cellular viability with no adverse effect on cell membrane depolarization. During depolarization of membrane potential, HEK 293T/17 cells labeled with the donor-acceptor FRET pair exhibit a greater fluorescence response in FluoVolt^™ emission relative to when FluoVolt^™ is used as the sole imaging probe. These results demonstrate the conjugated polyelectrolyte to be a new class of membrane labeling fluorophore for use in voltage sensing schemes.

Original language	English
Pages (from-to)	834-844
Number of pages	11
Journal	Photochemistry and Photobiology
Volume	96
Issue number	4
DOIs	https://doi.org/10.1111/php.13233
Publication status	Published - 2020 Jul 1

ASJC Scopus subject areas

Radiation
Biochemistry
Physical and Theoretical Chemistry

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10.1111/php.13233

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@article{f9f86dd474004d528478da462b14a88a,

title = "Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential",

abstract = "We report a F{\"o}rster resonance energy transfer (FRET)-based imaging ensemble for the visualization of membrane potential in living cells. A water-soluble poly(fluorene-cophenylene) conjugated polyelectrolyte (FsPFc10) serves as a FRET donor to a voltage-sensitive dye acceptor (FluoVolt{\texttrademark}). We observe FRET between FsPFc10 and FluoVolt{\texttrademark}, where the enhancement in FRET-sensitized emission from FluoVolt{\texttrademark} is measured at various donor/acceptor ratios. At a donor/acceptor ratio of 1, the excitation of FluoVolt{\texttrademark} in a FRET configuration results in a three-fold enhancement in its fluorescence emission (compared to when it is excited directly). FsPFc10 efficiently labels the plasma membrane of HEK 293T/17 cells and remains resident with minimal cellular internalization for ~ 1.5 h. The successful plasma membrane-associated colabeling of the cells with the FsPFc10-FluoVolt{\texttrademark} donor-acceptor pair is confirmed by dual-channel confocal imaging. Importantly, cells labeled with FsPFc10 show excellent cellular viability with no adverse effect on cell membrane depolarization. During depolarization of membrane potential, HEK 293T/17 cells labeled with the donor-acceptor FRET pair exhibit a greater fluorescence response in FluoVolt{\texttrademark} emission relative to when FluoVolt{\texttrademark} is used as the sole imaging probe. These results demonstrate the conjugated polyelectrolyte to be a new class of membrane labeling fluorophore for use in voltage sensing schemes.",

author = "Nag, {Okhil K.} and Jeong, {Ji Eun} and Le, {Van Sang} and Eunkeu Oh and Woo, {Han Young} and Delehanty, {James B.}",

note = "Funding Information: The authors acknowledge the NRL Base Funding Program and the NRL Institute for Nanoscience for financial support. H.Y.W. acknowledges the financial support from the National Research Foundation (NRF) of Korea (2019R1A2C2085290, 2019R1A6A1A11044070). Publisher Copyright: {\textcopyright} 2020 American Society for Photobiology",

year = "2020",

month = jul,

day = "1",

doi = "10.1111/php.13233",

language = "English",

volume = "96",

pages = "834--844",

journal = "Photochemistry and Photobiology",

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publisher = "Wiley-Blackwell",

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TY - JOUR

T1 - Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential

AU - Nag, Okhil K.

AU - Jeong, Ji Eun

AU - Le, Van Sang

AU - Oh, Eunkeu

AU - Woo, Han Young

AU - Delehanty, James B.

N1 - Funding Information: The authors acknowledge the NRL Base Funding Program and the NRL Institute for Nanoscience for financial support. H.Y.W. acknowledges the financial support from the National Research Foundation (NRF) of Korea (2019R1A2C2085290, 2019R1A6A1A11044070). Publisher Copyright: © 2020 American Society for Photobiology

PY - 2020/7/1

Y1 - 2020/7/1

N2 - We report a Förster resonance energy transfer (FRET)-based imaging ensemble for the visualization of membrane potential in living cells. A water-soluble poly(fluorene-cophenylene) conjugated polyelectrolyte (FsPFc10) serves as a FRET donor to a voltage-sensitive dye acceptor (FluoVolt™). We observe FRET between FsPFc10 and FluoVolt™, where the enhancement in FRET-sensitized emission from FluoVolt™ is measured at various donor/acceptor ratios. At a donor/acceptor ratio of 1, the excitation of FluoVolt™ in a FRET configuration results in a three-fold enhancement in its fluorescence emission (compared to when it is excited directly). FsPFc10 efficiently labels the plasma membrane of HEK 293T/17 cells and remains resident with minimal cellular internalization for ~ 1.5 h. The successful plasma membrane-associated colabeling of the cells with the FsPFc10-FluoVolt™ donor-acceptor pair is confirmed by dual-channel confocal imaging. Importantly, cells labeled with FsPFc10 show excellent cellular viability with no adverse effect on cell membrane depolarization. During depolarization of membrane potential, HEK 293T/17 cells labeled with the donor-acceptor FRET pair exhibit a greater fluorescence response in FluoVolt™ emission relative to when FluoVolt™ is used as the sole imaging probe. These results demonstrate the conjugated polyelectrolyte to be a new class of membrane labeling fluorophore for use in voltage sensing schemes.

AB - We report a Förster resonance energy transfer (FRET)-based imaging ensemble for the visualization of membrane potential in living cells. A water-soluble poly(fluorene-cophenylene) conjugated polyelectrolyte (FsPFc10) serves as a FRET donor to a voltage-sensitive dye acceptor (FluoVolt™). We observe FRET between FsPFc10 and FluoVolt™, where the enhancement in FRET-sensitized emission from FluoVolt™ is measured at various donor/acceptor ratios. At a donor/acceptor ratio of 1, the excitation of FluoVolt™ in a FRET configuration results in a three-fold enhancement in its fluorescence emission (compared to when it is excited directly). FsPFc10 efficiently labels the plasma membrane of HEK 293T/17 cells and remains resident with minimal cellular internalization for ~ 1.5 h. The successful plasma membrane-associated colabeling of the cells with the FsPFc10-FluoVolt™ donor-acceptor pair is confirmed by dual-channel confocal imaging. Importantly, cells labeled with FsPFc10 show excellent cellular viability with no adverse effect on cell membrane depolarization. During depolarization of membrane potential, HEK 293T/17 cells labeled with the donor-acceptor FRET pair exhibit a greater fluorescence response in FluoVolt™ emission relative to when FluoVolt™ is used as the sole imaging probe. These results demonstrate the conjugated polyelectrolyte to be a new class of membrane labeling fluorophore for use in voltage sensing schemes.

UR - http://www.scopus.com/inward/record.url?scp=85083794228&partnerID=8YFLogxK

U2 - 10.1111/php.13233

DO - 10.1111/php.13233

M3 - Article

C2 - 32083762

AN - SCOPUS:85083794228

SN - 0031-8655

VL - 96

SP - 834

EP - 844

JO - Photochemistry and Photobiology

JF - Photochemistry and Photobiology

IS - 4

ER -

Anionic Conjugated Polyelectrolytes for FRET-based Imaging of Cellular Membrane Potential

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