Two-photon fluorescence excitation cross sections of biomolecular probes from 690 to 960 nm

Albotǎ, Marius A.; Xu, Chris; Webb, Watt W.

doi:10.1364/ao.37.007352

Cited by 637 publications

(560 citation statements)

References 7 publications

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“…Although these Φ F σ 2 -values are lower than those of other organic fluorescent dyes (e.g. 10-100 GM 23 ), ANG-2 seems to be 2P-excitable. At 780 nm, the used 2P-excitation wavelength for in situ recordings in this study, an increase in Φ F σ 2 from (0.45 ± 0.01) GM for the Na + -free form to (5.91 ± 0.08) GM for the Na + -saturated form was observed.…”

Section: Photochemical and Photobiological Sciences Papermentioning

confidence: 58%

ANG-2 for quantitative Na+ determination in living cells by time-resolved fluorescence microscopy

Roder

Hille

2014

Photochem Photobiol Sci

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U n i v e r s i t ä t P o t s d a mPostprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe ; 184 Phillip Roder and Carsten Hille* Sodium ions (Na + ) play an important role in a plethora of cellular processes, which are complex and partly still unexplored. For the investigation of these processes and quantification of intracellular Na + concentrations ([Na + ] i ), two-photon coupled fluorescence lifetime imaging microscopy (2P-FLIM) was performed in the salivary glands of the cockroach Periplaneta americana. For this, the novel Na + -sensitive fluorescent dye Asante NaTRIUM Green-2 (ANG-2) was evaluated, both in vitro and in situ. In this context, absorption coefficients, fluorescence quantum yields and 2P action cross-sections were determined for the first time. ANG-2 was 2P-excitable over a broad spectral range and displayed fluorescence in the visible spectral range. Although the fluorescence decay behaviour of ANG-2 was triexponential in vitro, its analysis indicates a Na + -sensitivity appropriate for recordings in living cells. The Na + -sensitivity was reduced in situ, but the biexponential fluorescence decay behaviour could be successfully analysed in terms of quantitative [Na + ] i recordings. Thus, physiological 2P-FLIM measurements revealed a dopamineinduced [Na + ] i rise in cockroach salivary gland cells, which was dependent on a Na + -K + -2Cl − cotransporter (NKCC) activity. It was concluded that ANG-2 is a promising new sodium indicator applicable for diverse biological systems.

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Section: Photochemical and Photobiological Sciences Papermentioning

confidence: 58%

ANG-2 for quantitative Na+ determination in living cells by time-resolved fluorescence microscopy

Roder

Hille

2014

Photochem Photobiol Sci

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“…Two-photon absorption relies on the essentially instantaneous delivery of two low-energy photons to promote a chromophore from the ground state to the excited state (27), and is beneficial for imaging in thick tissue because lower-energy photons scatter less, cause less tissue damage, and, in the case of two-photon microscopy, reduce out-of-plane illumination because of the quadratic dependence of two-photon absorbance on incident light intensity (30). We measured the two-photon absorption cross-section (σ TPA ) of RVF5 and VF2.1.Cl in solution by normalizing to a rhodamine B standard (SI Appendix) (31)(32)(33). A plot of two-photon cross-section vs. excitation wavelength reveals a σ TPA maximum of 120 GM at 820 nm for RVF5 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Voltage-sensitive rhodol with enhanced two-photon brightness

Kulkarni

Kramer

Pourmandi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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We have designed, synthesized, and applied a rhodol-based chromophore to a molecular wire-based platform for voltage sensing to achieve fast, sensitive, and bright voltage sensing using twophoton (2P) illumination. Rhodol VoltageFluor-5 (RVF5) is a voltage-sensitive dye with improved 2P cross-section for use in thick tissue or brain samples. RVF5 features a dichlororhodol core with pyrrolidyl substitution at the nitrogen center. In mammalian cells under one-photon (1P) illumination, RVF5 demonstrates high voltage sensitivity (28% ΔF/F per 100 mV) and improved photostability relative to first-generation voltage sensors. This photostability enables multisite optical recordings from neurons lacking tuberous sclerosis complex 1, Tsc1, in a mouse model of genetic epilepsy. Using RVF5, we show that Tsc1 KO neurons exhibit increased activity relative to wild-type neurons and additionally show that the proportion of active neurons in the network increases with the loss of Tsc1. The high photostability and voltage sensitivity of RVF5 is recapitulated under 2P illumination. Finally, the ability to chemically tune the 2P absorption profile through the use of rhodol scaffolds affords the unique opportunity to image neuronal voltage changes in acutely prepared mouse brain slices using 2P illumination. Stimulation of the mouse hippocampus evoked spiking activity that was readily discerned with bath-applied RVF5, demonstrating the utility of RVF5 and molecular wire-based voltage sensors with 2P-optimized fluorophores for imaging voltage in intact brain tissue. Neuronal membrane potential dynamics drive neurotransmitter release and are therefore responsible for the unique physiology associated with neurons at cellular, circuit, and organismal levels. Despite the central importance of proper neuronal firing to human health, an integrated understanding of neuronal activity in the context of larger brain circuits remains elusive, due in part to a lack of methods for interrogating membrane potential dynamics with sufficient spatial and temporal resolution.Traditional methods for monitoring membrane potential rely heavily on the use of invasive electrodes, through one of two methods. The first method, patch-clamp electrophysiology, uses a single electrode to make contact with or puncture a cell to record changes in membrane potential, sacrificing throughput and spatial resolution to achieve a comprehensive description of a single cellular electrophysiological profile. A second method uses multielectrode arrays (MEAs), in which patterned arrays of electrodes introduced to cells or tissues report on electrical changes. Spatial resolution of MEAs depends on the number and positioning of the electrodes within the array. Although throughput is improved relative to patch-clamp electrophysiology, the extracellularly recorded signals are typically less sensitive than whole-cell methods and can be an amalgamation of several cells, making deconvolution of recorded signals and precise correlation to specific cells difficult or impossible. Addi...

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“…TPEF measurements were calibrated relative to the absolute TPEF action crosssection determined by Xu and Webb for fluorescein in water (pH ) 11) in the 690-1000-nm range. 90,91 For each data point, an additional control was performed using the known TPEF action cross-section of rhodamine B in methanol. 90, 91 The experimental uncertainty amounts to (10%.…”

Section: Scheme 1: Synthesis Of Chromophores 1-3mentioning

confidence: 99%

“…90,91 For each data point, an additional control was performed using the known TPEF action cross-section of rhodamine B in methanol. 90, 91 The experimental uncertainty amounts to (10%.…”

Section: Scheme 1: Synthesis Of Chromophores 1-3mentioning

confidence: 99%

Effects of (Multi)branching of Dipolar Chromophores on Photophysical Properties and Two-Photon Absorption

et al. 2005

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To investigate the effect of branching on linear and nonlinear optical properties, a specific series of chromophores, epitome of (multi)branched dipoles, has been thoroughly explored by a combined theoretical and experimental approach. Excited-state structure calculations based on quantum-chemical techniques (timedependent density functional theory) as well as a Frenkel exciton model nicely complement experimental photoluminescence and one-and two-photon absorption findings and contribute to their interpretation. This allowed us to get a deep insight into the nature of fundamental excited-state dynamics and the nonlinear optical (NLO) response involved. Both experiment and theory reveal that a multidimensional intramolecular charge transfer takes place from the donating moiety to the periphery of the branched molecules upon excitation, while fluorescence stems from an excited state localized on one of the dipolar branches. Branching is also observed to lead to cooperative enhancement of two-photon absorption (TPA) while maintaining high fluorescence quantum yield, thanks to localization of the emitting state. The comparison between results obtained in the Frenkel exciton scheme and ab initio results suggests the coherent coupling between branches as one of the possible mechanisms for the observed enhancement. New strategies for the rational design of NLO molecular assemblies are thus inferred on the basis of the acquired insights.

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Two-photon fluorescence excitation cross sections of biomolecular probes from 690 to 960 nm

Cited by 637 publications

References 7 publications

ANG-2 for quantitative Na+ determination in living cells by time-resolved fluorescence microscopy

ANG-2 for quantitative Na+ determination in living cells by time-resolved fluorescence microscopy

Voltage-sensitive rhodol with enhanced two-photon brightness

Effects of (Multi)branching of Dipolar Chromophores on Photophysical Properties and Two-Photon Absorption

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