Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye

Bright biocompatible fluorescent imaging dyes with red to near-infrared (NIR) emissions are ideal candidates for fluorescence microscopy applications. Pyrene–benzothiazolium hemicyanine dyes are a new class of lysosome-specific probes reported on recently. In this work, we conduct a detailed impleme...

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Main Authors: Chathura S. Abeywickrama, Enya Huang, Wenhui Yan, Michael A. Vrionides, Paaramitha Warushavithana, Kristen A. Johnson, Robert V. Stahelin, Yi Pang, Tomoyasu Mani, Kaveesha J. Wijesinghe
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Language:English
Published: MDPI AG 2024-12-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/14/12/612
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author Chathura S. Abeywickrama
Enya Huang
Wenhui Yan
Michael A. Vrionides
Paaramitha Warushavithana
Kristen A. Johnson
Robert V. Stahelin
Yi Pang
Tomoyasu Mani
Kaveesha J. Wijesinghe
author_facet Chathura S. Abeywickrama
Enya Huang
Wenhui Yan
Michael A. Vrionides
Paaramitha Warushavithana
Kristen A. Johnson
Robert V. Stahelin
Yi Pang
Tomoyasu Mani
Kaveesha J. Wijesinghe
author_sort Chathura S. Abeywickrama
collection DOAJ
description Bright biocompatible fluorescent imaging dyes with red to near-infrared (NIR) emissions are ideal candidates for fluorescence microscopy applications. Pyrene–benzothiazolium hemicyanine dyes are a new class of lysosome-specific probes reported on recently. In this work, we conduct a detailed implementation study for a pyrene–benzothiazolium derivative, BTP, to explore its potential imaging applications in fluorescence microscopy. The optical properties of BTP are studied in intracellular environments through advanced fluorescence microscopy techniques, with BTP exhibiting a noticeable shift toward blue (λ<sub>em</sub> ≈ 590 nm) emissions in cellular lysosomes. The averaged photon arrival time (AAT)-based studies exhibit two different emissive populations of photons, indicating the probe’s dynamic equilibrium between two distinctively different lysosomal microenvironments. Here, BTP is successfully utilized for time-lapse fluorescence microscopy imaging in real-time as a ‘wash-free’ imaging dye with no observed background interference. BTP exhibits an excellent ability to highlight microorganisms (i.e., bacteria) such as <i>Bacillus megaterium</i> through fluorescence microscopy. BTP is found to be a promising candidate for two-photon fluorescence microscopy imaging. The two-photon excitability of BTP in COS-7 cells is studied, with the probe exhibiting an excitation maximum at λ<sub>TP</sub> ≈ 905 nm.
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spelling doaj-art-7de1709485014fdd8e0584eefdfaa08f2025-08-20T02:00:51ZengMDPI AGBiosensors2079-63742024-12-01141261210.3390/bios14120612Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium DyeChathura S. Abeywickrama0Enya Huang1Wenhui Yan2Michael A. Vrionides3Paaramitha Warushavithana4Kristen A. Johnson5Robert V. Stahelin6Yi Pang7Tomoyasu Mani8Kaveesha J. Wijesinghe9Department of Pharmaceutical Sciences, University of Connecticut, Storrs, CT 06269, USADepartment of Pharmaceutical Sciences, University of Connecticut, Storrs, CT 06269, USADepartment of Pharmaceutical Sciences, University of Connecticut, Storrs, CT 06269, USADepartment of Chemistry, University of Connecticut, Storrs, CT 06269, USADepartment of Chemistry, University of Colombo, Colombo 00300, Sri LankaDepartment of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USABorch Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USADepartment of Chemistry, The University of Akron, Akron, OH 44325, USADepartment of Chemistry, University of Connecticut, Storrs, CT 06269, USADepartment of Chemistry, University of Colombo, Colombo 00300, Sri LankaBright biocompatible fluorescent imaging dyes with red to near-infrared (NIR) emissions are ideal candidates for fluorescence microscopy applications. Pyrene–benzothiazolium hemicyanine dyes are a new class of lysosome-specific probes reported on recently. In this work, we conduct a detailed implementation study for a pyrene–benzothiazolium derivative, BTP, to explore its potential imaging applications in fluorescence microscopy. The optical properties of BTP are studied in intracellular environments through advanced fluorescence microscopy techniques, with BTP exhibiting a noticeable shift toward blue (λ<sub>em</sub> ≈ 590 nm) emissions in cellular lysosomes. The averaged photon arrival time (AAT)-based studies exhibit two different emissive populations of photons, indicating the probe’s dynamic equilibrium between two distinctively different lysosomal microenvironments. Here, BTP is successfully utilized for time-lapse fluorescence microscopy imaging in real-time as a ‘wash-free’ imaging dye with no observed background interference. BTP exhibits an excellent ability to highlight microorganisms (i.e., bacteria) such as <i>Bacillus megaterium</i> through fluorescence microscopy. BTP is found to be a promising candidate for two-photon fluorescence microscopy imaging. The two-photon excitability of BTP in COS-7 cells is studied, with the probe exhibiting an excitation maximum at λ<sub>TP</sub> ≈ 905 nm.https://www.mdpi.com/2079-6374/14/12/612pyrene–benzothiazoliumfluorescence microscopycell imagingfluorescence lifetimelysosomeStokes’ shift
spellingShingle Chathura S. Abeywickrama
Enya Huang
Wenhui Yan
Michael A. Vrionides
Paaramitha Warushavithana
Kristen A. Johnson
Robert V. Stahelin
Yi Pang
Tomoyasu Mani
Kaveesha J. Wijesinghe
Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
Biosensors
pyrene–benzothiazolium
fluorescence microscopy
cell imaging
fluorescence lifetime
lysosome
Stokes’ shift
title Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
title_full Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
title_fullStr Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
title_full_unstemmed Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
title_short Exploring Imaging Applications of a Red-Emitting π-Acceptor (π-A) Pyrene-Benzothiazolium Dye
title_sort exploring imaging applications of a red emitting π acceptor π a pyrene benzothiazolium dye
topic pyrene–benzothiazolium
fluorescence microscopy
cell imaging
fluorescence lifetime
lysosome
Stokes’ shift
url https://www.mdpi.com/2079-6374/14/12/612
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