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Label-Free Ratiometric Imaging of Serotonin in Live Cells
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    Label-Free Ratiometric Imaging of Serotonin in Live Cells
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    Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), Homi Bhabha Road, Colaba, Mumbai 400005, India
    Department of Applied Physics, Indian Institute of Technology (Indian School of Mines), Dhanbad, 826004 Jharkhand, India
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    ACS Chemical Neuroscience

    Cite this: ACS Chem. Neurosci. 2017, 8, 11, 2369–2373
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    https://doi.org/10.1021/acschemneuro.7b00132
    Published August 10, 2017
    Copyright © 2017 American Chemical Society

    Abstract

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    Ratiometric imaging can quantitatively measure changes in cellular analyte concentrations using specially designed fluorescent labels. We describe a label-free ratiometric imaging technique for direct detection of changes in intravesicular serotonin concentration in live cells. At higher concentrations, serotonin forms transient oligomers whose ultraviolet emission is shifted to longer wavelengths. We access the ultraviolet/blue emission using relatively benign three-photon excitation and split it into two imaging channels, whose ratio reports the concentration. The technique is sensitive at a physiologically relevant concentration range (10–150 mM serotonin). As a proof of principle, we measure the increase of intravesicular serotonin concentration with the addition of external serotonin. In general, since emission spectra of molecules are often sensitive to concentration, our method may be applicable to other natively fluorescent intracellular molecules which are present at high concentrations.

    Copyright © 2017 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acschemneuro.7b00132.

    • Graph of fluorescence signal vs excitation power indicating three-photon excitation, serotonin fluorescence vs pH, and effect of SERT blocker on serotonin uptake (PDF)

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    Cited By

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    This article is cited by 13 publications.

    1. Kallol Bera, Anand Kant Das, Ananya Rakshit, Bidyut Sarkar, Anoop Rawat, Barun Kumar Maity, Sudipta Maiti. Fluorogenic Detection of Monoamine Neurotransmitters in Live Cells. ACS Chemical Neuroscience 2018, 9 (3) , 469-474. https://doi.org/10.1021/acschemneuro.7b00391
    2. Megha Maheshwari, Aastha Singla, Anoop Rawat, Toshali Banerjee, Sthitapranjya Pati, Sneha Shah, Sudipta Maiti, Vidita A. Vaidya. Chronic chemogenetic activation of hippocampal progenitors enhances adult neurogenesis and modulates anxiety-like behavior and fear extinction learning. IBRO Neuroscience Reports 2024, 16 , 168-181. https://doi.org/10.1016/j.ibneur.2024.01.002
    3. Barun Kumar Maity, Debsankar Saha Roy, Sudipta Maiti. Real time imaging of the excitation volume of a multiphoton microscope. Journal of Optics 2022, 24 (6) , 064012. https://doi.org/10.1088/2040-8986/ac69f5
    4. Sakshi, Bikash Chandra Swain, Anand Kant Das, Nitesh Kumar Pathak, Umakanta Tripathy. Z-scan analysis and theoretical studies of dopamine under physiological conditions. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 2022, 271 , 120890. https://doi.org/10.1016/j.saa.2022.120890
    5. Yasuaki Kumamoto. Deep-ultraviolet microscopy for tryptophan label-free imaging in cells and tissue. 2022, 25-39. https://doi.org/10.1016/B978-0-12-822790-9.00003-6
    6. Sakshi, Bikash Chandra Swain, Anand Kant Das, Nitesh Kumar Pathak, Umakanta Tripathy. Norepinephrine exhibits thermo-optical nonlinearity under physiological conditions. Physical Chemistry Chemical Physics 2021, 23 (41) , 23473-23477. https://doi.org/10.1039/D1CP03534G
    7. Guillermo A. Hernández-Mendoza, Dilia Aguirre-Olivas, Mario González-Gutiérrez, Héctor J. Leal, Naser Qureshi, Carlos G. Treviño-Palacios, Jorge Peón, Francisco F. De-Miguel. Fluorescence of serotonin in the visible spectrum upon multiphotonic photoconversion. Biomedical Optics Express 2020, 11 (3) , 1432. https://doi.org/10.1364/BOE.380412
    8. Sakshi, Bikash Chandra Swain, Anand Kant Das, Umakanta Tripathy. Probing third-order nonlinearity in serotonin: A Z-scan study. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 2019, 223 , 117319. https://doi.org/10.1016/j.saa.2019.117319
    9. Sthitapranjya Pati, Sonali S. Salvi, Mamata Kallianpur, Bhupesh Vaidya, Antara Banerjee, Sudipta Maiti, James P. Clement, Vidita A. Vaidya. Chemogenetic Activation of Excitatory Neurons Alters Hippocampal Neurotransmission in a Dose-Dependent Manner. eneuro 2019, 6 (6) , ENEURO.0124-19.2019. https://doi.org/10.1523/ENEURO.0124-19.2019
    10. Yasuaki Kumamoto, Atsushi Taguchi, Satoshi Kawata. Deep‐Ultraviolet Biomolecular Imaging and Analysis. Advanced Optical Materials 2019, 7 (5) https://doi.org/10.1002/adom.201801099
    11. Shrutidhara Biswas, Vlad Bogdan Gavra, Anand Kant Das, Umakanta Tripathy. Biophotonics in Disease Diagnosis and Therapy. 2019, 65-86. https://doi.org/10.1007/978-981-13-3705-5_3
    12. Parijat Sarkar, G. Aditya Kumar, Sreetama Pal, Amitabha Chattopadhyay. Biophysics of Serotonin and the Serotonin1A Receptor. 2019, 3-22. https://doi.org/10.1016/B978-0-12-800050-2.00001-2
    13. Barun Kumar Maity, Sudipta Maiti. Label-free imaging of neurotransmitters in live brain tissue by multi-photon ultraviolet microscopy. Neuronal Signaling 2018, 2 (4) https://doi.org/10.1042/NS20180132

    ACS Chemical Neuroscience

    Cite this: ACS Chem. Neurosci. 2017, 8, 11, 2369–2373
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acschemneuro.7b00132
    Published August 10, 2017
    Copyright © 2017 American Chemical Society

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