Ultrafast supercontinuum sculpting for two-photon spectroscopy and microscopy of ratiometric fluorescent indicators

被引:1
作者
Chebotarev, Artem S. [1 ]
Raevsky, Roman I. [2 ,3 ]
Linovsky, Georgy N. [1 ]
Kostyuk, Alexander I. [2 ,3 ]
Belousov, Vsevolod V. [2 ,3 ,4 ]
Fedotov, Andrei B. [1 ]
Bilan, Dmitry S. [2 ,3 ]
Lanin, Aleksandr A. [1 ]
机构
[1] Moscow MV Lomonosov State Univ, Phys Dept, Moscow 111992, Russia
[2] Russian Acad Sci, MM Shemyakin & Yu A Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia
[3] Pirogov Russian Natl Res Med Univ, Moscow 117997, Russia
[4] Fed Ctr Brain Res & Neurotechnol, Fed Med Biol Agcy, Moscow 119330, Russia
基金
俄罗斯科学基金会;
关键词
HYDROGEN-PEROXIDE; ABSORPTION; H2O2; STANDARDS; PROBE;
D O I
10.1063/5.0197580
中图分类号
O59 [应用物理学];
学科分类号
摘要
We present a compact laser system for quantitative two-photon excitation spectra measurements and ratiometric two-photon imaging of fluorescent protein indicators. The fundamental of the system is a short segment of photonic crystal fiber (PCF), which supports a nonlinear transformation of low-power ultrashort pulses by preserving temporal coherence, and this generates an ultrafast almost octave-spanning supercontinuum (SC). Accurate sculpting of the SC by its amplitude and phase modulation provides implementation of the spectroscopic and microscopic modalities. The spectroscopic one was exhibited by two-photon action cross section spectra measuring for the genetically encoded fluorescent sensing proteins of the vital biochemical parameters: acidity (SypHer3s), concentration of hydrogen peroxide (HyPer3 and HyPer7), redox status of NADH and glutathione (RexYFP and Grx1-roGFP2), hypohalous acids and their derivatives (Hypocrates). For the microscopy, we investigated and optimized the intensity pump pulse profiles under the high numerical objective by dispersion scan technique. We conducted real-time monitoring of the dynamics of hydrogen peroxide in HeLa cells with subcellular spatial resolution by means of ratiometric two-photon imaging of Hyper7 sensors. The presented hybrid laser system provides an ideal optical toolbox in order to develop ratiometric fluorescent sensors, which can be visualized in vivo using two-photon microscopy.
引用
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页数:7
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共 49 条
  • [1] Self-calibrating d-scan: measuring ultrashort laser pulses on-target using an arbitrary pulse compressor
    Alonso, Benjamin
    Sola, Inigo J.
    Crespo, Helder
    [J]. SCIENTIFIC REPORTS, 2018, 8
  • [2] Impact of the Protein Environment on Two-Photon Absorption Cross-Sections of the GFP Chromophore Anion Resolved at the XMCQDPT2 Level of Theory
    Aslopovsky, Vladislav R.
    Scherbinin, Andrei V.
    Kleshchina, Nadezhda N.
    Bochenkova, Anastasia V.
    [J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2023, 24 (14)
  • [3] Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence
    Barnett, Lauren M.
    Hughes, Thomas E.
    Drobizhev, Mikhail
    [J]. PLOS ONE, 2017, 12 (02):
  • [4] In Vivo Imaging of Hydrogen Peroxide with HyPer Probes
    Bilan, Dmitry S.
    Belousov, Vsevolod V.
    [J]. ANTIOXIDANTS & REDOX SIGNALING, 2018, 29 (06) : 569 - 584
  • [5] Genetically encoded fluorescent indicator for imaging NAD+/NADH ratio changes in different cellular compartments
    Bilan, Dmitry S.
    Matlashov, Mikhail E.
    Gorokhovatsky, Andrey Yu.
    Schultz, Carsten
    Enikolopov, Grigori
    Belousov, Vsevolod V.
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2014, 1840 (03): : 951 - 957
  • [6] HyPer-3: A Genetically Encoded H2O2 Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging
    Bilan, Dmitry S.
    Pase, Luke
    Joosen, Linda
    Gorokhovatsky, Andrey Yu
    Ermakova, Yulia G.
    Gadella, Theodorus W. J.
    Grabher, Clemens
    Schultz, Carsten
    Lukyanov, Sergey
    Belousov, Vsevolod V.
    [J]. ACS CHEMICAL BIOLOGY, 2013, 8 (03) : 535 - 542
  • [7] Multiparametric optical analysis of mitochondrial redox signals during neuronal physiology and pathology in vivo
    Breckwoldt, Michael
    Pfister, Franz M. J.
    Bradley, Peter M.
    Marinkovic, Petar
    Williams, Philip R.
    Brilll, Monika S.
    Plomer, Barbara
    Schmalz, Anja
    St Clair, Daret K.
    Naumann, Ronald
    Griesbeck, Oliver
    Schwarzlaender, Markus
    Godinho, Leanne
    Bareyre, Florence M.
    Dick, Tobias P.
    Kerschensteiner, Martin
    Misgeld, Thomas
    [J]. NATURE MEDICINE, 2014, 20 (05) : 559 - 122
  • [8] SUPEROXIDE AND HYDROGEN-PEROXIDE IN RELATION TO MAMMALIAN-CELL PROLIFERATION
    BURDON, RH
    [J]. FREE RADICAL BIOLOGY AND MEDICINE, 1995, 18 (04) : 775 - 794
  • [9] Multiphoton tools for hydrogen peroxide imaging in vivo with subcellular resolution
    Chebotarev, Artem S.
    V. Kelmanson, Ilya
    Ivanova, Alexandra D.
    V. Khramova, Yulia
    Katrukha, Veronika A.
    Kotova, Daria A.
    Raevskii, Roman I.
    Moshchenko, Aleksandr A.
    Linovsky, Georgy N.
    Fedotov, Andrei B.
    V. Belousov, Vsevolod
    Bilan, Dmitry S.
    Lanin, Aleksandr A.
    [J]. SENSORS AND ACTUATORS B-CHEMICAL, 2024, 410
  • [10] Single-beam dual-color alternate-pathway two-photon spectroscopy: Toward an optical toolbox for redox biology
    Chebotarev, Artem S.
    Lanin, Aleksandr A.
    Raevskii, Roman, I
    Kostyuk, Alexander, I
    Smolyarova, Daria D.
    Bilan, Dmitry S.
    Savitskii, Ilya, V
    Fedotov, Andrei B.
    Belousov, Vsevolod V.
    Zheltikov, Aleksei M.
    [J]. JOURNAL OF RAMAN SPECTROSCOPY, 2021, 52 (09) : 1552 - 1560