Gas-phase Fluorescence Research Articles

Fluorescence of BODIPY dyes in gas phase at near-ambient conditions

Molecular fluorescence is a phenomenon that is usually observed in condensed phase. It is strongly affected by molecular interactions. The study of fluorescence spectra in gas-phase can provide a nearly ideal model for the evaluation of the intrinsic properties of fluorophores. Unfortunately, most conventional fluorophores are not volatile enough to allow for the study of their fluorescence in gas phase. Here we report very bright gas phase fluorescence of simple BODIPY dyes that can be readily observed at atmospheric pressure using conventional fluorescence instrumentation. To our knowledge, this is the first example of visible-range gas-phase fluorescence at near-ambient conditions. Evaporation of the dye in vacuum allowed us to demonstrate organic molecular electroluminescence in gas discharge excited by the electric field produced by a Tesla coil.

Empirical Calibration of a Cylindrical Ion Trap for Mass-Selected Gas-Phase Fluorescence Spectroscopy.

The ion motion in a quadrupole ion trap of hyperbolic geometry is well described by the Mathieu equations. A simpler cylindrical ion trap has also gained significance and has been used by us for fluorescence-spectroscopy experiments. This design allows for the easy replacement of the end-cap with a mesh, enhancing the photon collection. It is crucial to obtain a firm understanding of the ion motion in cylindrical ion traps and their capability as mass spectrometers. We present here an empirical method of calibrating a cylindrical ion trap based on fluorescence detection. This can be done nearly background-free in a pulsed experiment. The ions are located at the center of the trap, where the field is primarily quadrupolar, and here an effective Mathieu description is found through an effective geometry parameter. In spectroscopy experiments, high buffer-gas pressures are needed to efficiently cool the ions, which complicates the ions' motion and hence their stability. Still, simulations show that the stability diagram closely aligns with the Mathieu diagram, albeit shifted due to collisions. We map the stability diagram for six molecular ions by fluorescence collection from four cations and two anions spanning m/z from 212 to 647. The stability diagram is parametrized through the Mathieu functions with an m/z-dependent effective geometry parameter and a q-dependent shrinkage of the diagram. Based on the calibration, we estimate the mass resolution to be +7/-3 Da for ions with masses in the hundreds of Da.

Cryogenic Fluorescence Spectroscopy of Ionic Fluorones in Gaseous and Condensed Phases: New Light on Their Intrinsic Photophysics.

Fluorescence spectroscopy of gas-phase ions generated through electrospray ionization is an emerging technique able to probe intrinsic molecular photophysics directly without perturbations from solvent interactions. While there is ample scope for the ongoing development of gas-phase fluorescence techniques, the recent expansion into low-temperature operating conditions accesses a wealth of data on intrinsic fluorophore photophysics, offering enhanced spectral resolution compared with room-temperature measurements, without matrix effects hindering the excited-state dynamics. This perspective reviews current progress on understanding the photophysics of anionic fluorone dyes, which exhibit an unusually large Stokes shift in the gas phase, and discusses how comparison of gas- and condensed-phase fluorescence spectra can fingerprint structural dynamics. The capacity for temperature-dependent measurements of both fluorescence emission and excitation spectra helps establish the foundation for the use of fluorone dyes as fluorescent tags in macromolecular structure determination. We suggest ideas for technique development.

Laser-Induced Fluorescence (LIF) Spectroscopy of Trapped Molecular Ions in the Gas Phase.

This review focuses on the laser-induced fluorescence (LIF) spectroscopy of trapped gas-phase molecular ions, a developing field of research. Following a brief description of the theory and experimental approaches employed in general for fluorescence spectroscopy, the review summarizes the current state-of-the-art intrinsic fluorescence measurement techniques employed for gas-phase ions. Whereas the LIF spectroscopy of condensed matter systems is a well-developed area of research, the instrumentation used for such studies is not directly applicable to gas-phase ions. However, some measurement schemes employed in condensed-phase experiments could be highly beneficial for gas-phase investigations. We have included a brief discussion on some of these techniques as well. Quadrupole ion traps are commonly used for spatial confinement of ions in the ion-trap-based LIF. One of the main challenges involved in such experiments is the poor signal-to-noise ratio (SNR) arising due to weak gas-phase fluorescence emission, high background noise, and small solid angle for the fluorescence collection optics. The experimental approaches based on the integrated high-finesse optical cavities employed for the condensed-phase measurements provide a better (typically an order of magnitude more) SNR in the detected fluorescence than the single-pass detection schemes. Another key to improving the SNR is to exploit the maximum solid angle of light collection by choosing high numerical aperture (NA) collection optics. A combination of these two approaches integrated with ion traps could transmogrify this field, allowing one to study even weak fluorescence emission from gas-phase molecular ions. The review concludes by discussing the scope of the advances in the LIF instrumentation for detailed spectral characterization of fluorophores of weak gas-phase fluorescence emission, considering fluorescein as one example.

Gas-Phase Fluorescence of Proflavine Reveals Two Close-Lying, Brightly Emitting States.

Surprising excitation-dependent, dual emission from a small organic model fluorophore is reported. Gas-phase fluorescence spectra of proflavine (a diaminoacridine) ions reveal two long-lived emitting states, with distinct bands separated by just 1700 cm-1. The relative intensities of these two bands depend on the excitation wavelength. Time-dependent density functional theory (TD-DFT) calculations support the existence of two close-lying singlet electronic states, with excitation into S2 predicted to be >1000-fold more likely than into S1. These data strongly suggest that internal conversion (IC) rates are suppressed relative to solvated proflavine, and that IC is competitive with intramolecular vibrational relaxation (IVR). This work offers an in-depth assessment of the gas-phase photophysics of a simple fluorophore that could open a new pathway to understanding dual emission in fluorophores.

Intrinsic fluorescence from firefly oxyluciferin monoanions isolated in vacuo.

Fireflies, click beetles, and railroad worms glow in the dark. The color varies from green to red among the insects and is associated with an electronically excited oxyluciferin formed catalytically by the luciferase enzyme. The actual color tuning mechanism has been, and still is, up for much debate. One complication is that oxyluciferin can occur in different charge states and isomeric forms. We present here emission spectra of oxyluciferin monoanions in vacuo at both room temperature and at 100 K recorded with a newly developed and unique mass-spectroscopy setup specially designed for gas-phase ion fluorescence spectroscopy. Ions are limited to the phenolate-keto and phenolate-enol forms that account for natural bioluminescence. At 100 K, fluorescence band maxima are at 599 ± 2 nm and 563 ± 2 nm for the keto and enol forms, respectively, and at 300 K about 5 nm further to the red. The bare-ion spectra, free from solvent effects, serve as important references as they reveal whether a protein microenvironment redshifts or blueshifts the emission, and they serve as important benchmarks for nontrivial excited-state calculations.

Adapting a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer for Gas-Phase Fluorescence Spectroscopy Measurement of Trapped Biomolecular Ions.

Gas-phase fluorescence spectroscopy is still in its infancy, which demands further instrumental developments. In this study, a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS), equipped with a lab-developed data acquisition system, was coupled to a tunable femtosecond laser and a state-of-the-art optical system for fluorescence studies of mass-selected ions. For excitation, a laser beam was focused (beam size < 1.0 mm) into the cylindrical ICR cell. A wire mesh replaced the back trapping plate, allowing ∼10% of the fluorescence emitted from trapped ions to be collected by a lens installed beside the wire mesh. The collected fluorescence light was then transmitted outside of the mass spectrometer via fiber optics. A novel accumulation during detection (ADD) scheme was developed to increase the duty cycle of gas-phase fluorescence spectroscopy experiments. With ADD, >90% duty cycle for mass spectrometry and fluorescence experiments could be achieved. This instrument was able to perform fluorescence experiments on various ions, from simple rhodamine dyes to large biomolecules (i.e., peptides and proteins) labeled with dyes of various optical properties. A fluorescence lifetime measurement of trapped rhodamine 6G cations was also performed, yielding a value of 5.97 ± 0.23 ns. This setup has a broad mass range and decent fluorescence spectroscopy performance (i.e., the emission spectrum of rhodamine 6G can be acquired with good S/N in a minute). Finally, this setup also allows more challenging gas-phase fluorescence spectroscopy experiments, for example, of low quantum yield fluorophores and large biomolecules in their native state that appear at high m/z, which may not be doable with quadrupole ion traps (QIT).

Effect of Freezing out Vibrational Modes on Gas-Phase Fluorescence Spectra of Small Ionic Dyes.

While action spectroscopy of cold molecular ions is a well-established technique to provide vibrationally resolved absorption features, fluorescence experiments are still challenging. Here we report the fluorescence spectra of pyronin-Y and resorufin ions at 100 K using a newly constructed setup. Spectra narrow upon cooling, and the emission maxima blueshift. Temperature effects are attributed to the population of vibrational excited levels in S1, and that frequencies are lower in S1 than in S0. This picture is supported by calculated spectra based on a Franck-Condon model that not only predicts the observed change in maximum, but also assigns Franck-Condon active vibrations. In-plane vibrational modes that preserve the mirror plane present in both S0 and S1 of resorufin and pyronin Y account for most of the observed vibrational bands. Finally, at low temperatures, it is important to pick an excitation wavelength as far to the red as possible to not reheat the ions.

Fluorescence-based Techniques to Study the Structure and Dynamics of Mass-selected Biomolecular Ions.

Laser-induced fluorescence studies on mass-selected biomolecules are a promising route to understand their properties in the gas phase and probe their intrinsic properties in a solvent-free environment. Fluorescence has been used to investigate the conformation and dynamics of gaseous biomolecular ions. With Förster Resonance Energy Transfer (FRET), it is now possible to obtain sensitive intramolecular distance information from large biomolecules, like proteins, with high chemical specificity. With growing interest and applications, gas-phase fluorescence measurements can shed greater light on the characteristics of proteins in the gas phase. Compared to the solution phase measurements, gas-phase fluorescence can also help understand the influence of solvent interactions on the protein structure and function.

Fluorescence-Based Detection of the Desolvation Process of Protein Ions Generated in an Aqueous Electrospray Plume.

A new experimental setup to study laser-induced fluorescence from analytes at different locations in an electrospray plume has been developed. The high fluorescence collection efficiency (∼2%) of the setup, along with a sensitive charge coupled device (CCD) detector, enabled the study of low ion concentrations (down to ∼fM) in the plume. The use of small electrospray tip inner diameters (<1 μm) facilitated the fast desolvation of gaseous protein ions in an aqueous electrospray plume. Fluorescence spectra were acquired from specific locations along the plume axis in different aqueous electrospray plumes with three different analytes: a rhodamine dye and two proteins (ubiquitin and apomyoglobin) labeled with rhodamine dyes. To confirm the presence of gaseous ions, pure gas-phase fluorescence spectra were acquired in the vacuum of a modified ion trap mass spectrometer. These spectra were used to fit to confirm the presence of gaseous species in the corresponding spectra obtained from the electrospray plume. This study shows that with small inner diameter spray capillaries, gaseous protein ions generated at atmospheric pressure in an electrospray plume can be detected with fluorescence-based techniques. Fluorescence measurements can be used to study their structure in the electrospray plume, and the dynamics as they transition from solution to the gas phase and in the early stages after desolvation from charged droplets. Other techniques can also be applied to further study gaseous biomolecular structures under ambient conditions immediately after desolvation.

Photophysics of Isolated Rose Bengal Anions

Dye molecules based on the xanthene moiety are widely used as fluorescent probes in bioimaging and technological applications due to their large absorption cross-section for visible light and high fluorescence quantum yield. These applications require a clear understanding of the dye's inherent photophysics and the effect of a condensed-phase environment. Here, the gas-phase photophysics of the rose bengal doubly deprotonated dianion [RB - 2H]2-, deprotonated monoanion [RB - H]-, and doubly deprotonated radical anion [RB - 2H]•- is investigated using photodetachment, photoelectron, and dispersed fluorescence action spectroscopies, and tandem ion mobility spectrometry (IMS) coupled with laser excitation. For [RB - 2H]2-, photodetachment action spectroscopy reveals a clear band in the visible (450-580 nm) with vibronic structure. Electron affinity and repulsive Coulomb barrier (RCB) properties of the dianion are characterized using frequency-resolved photoelectron spectroscopy, revealing a decreased RCB compared with that of fluorescein dianions due to electron delocalization over halogen atoms. Monoanions [RB - H]- and [RB - 2H]•- differ in nominal mass by 1 Da but are difficult to study individually using action spectroscopies that isolate target ions using low-resolution mass spectrometry. This work shows that the two monoanions are readily distinguished and probed using the IMS-photo-IMS and photo-IMS-photo-IMS strategies, providing distinct but overlapping photodissociation action spectra in the visible spectral range. Gas-phase fluorescence was not detected from photoexcited [RB - 2H]2- due to rapid electron ejection. However, both [RB - H]- and [RB - 2H]•- show a weak fluorescence signal. The [RB - H]- action spectra show a large Stokes shift of ∼1700 cm-1, while the [RB - 2H]•- action spectra show no appreciable Stokes shift. This difference is explained by considering geometries of the ground and fluorescing states.

Gas-phase action and fluorescence spectroscopy of mass-selected fluorescein monoanions and two derivatives.

Gaseous fluorescein monoanions are weakly fluorescent; they display a broad fluorescence spectrum and a large Stokes shift. This contrasts with the situation in aqueous solution. One explanation of the intriguing behavior in vacuo is based on internal proton transfer from the pendant carboxyphenyl group to one of the xanthene oxygens in the excited state; another that rotation of the carboxyphenyl group relative to the xanthene leads to a partial charge transfer from one chromophore (xanthene) to the other (carboxyphenyl) when the π orbitals start to overlap. To shed light on the mechanism at play, we synthesized two fluorescein derivatives where the carboxylic acid group is replaced with either an ester or a tertiary amide functionality and explored their gas-phase ion fluorescence using the home-built LUminescence iNstrument in Aarhus (LUNA) setup. Results on the fluorescein methyl ester that has no acidic proton clearly disprove the former explanation: The spectrum remains broad, and the band center (at 605 nm) is shifted even more to the red than that of fluorescein (590 nm). Experiments on the other variant that contains a piperidino amide are also in favor of the second explanation as here the piperidino already causes the dihedral angle between the planes defining the xanthene and the benzene ring to be less than 90° in the ground state (i.e., 63°), according to density functional theory calculations. As a result of the closer similarity between the ground-state and excited-state structures, the fluorescence spectrum is narrower than those of the other two ions, and the band maximum is further to the blue (575 nm). In accordance with a more delocalized ground state of the amide derivative, action spectra associated with photoinduced dissociation recorded at another setup show that the absorption-band maximum for the amide derivative is redshifted compared to that of fluorescein (538 nm vs. 525 nm).

Carbazole and fluorene polyaniline derivatives: Synthesis, properties and application as multiple stimuli-responsive fluorescent chemosensor

Development of conjugated polymers with fluorescence sensing characteristics has received close attention from researchers in fields of environmental protection, biosensing and toxins detection on food. In this paper, novel polyaniline derivatives of poly(9-methyl-9H-carbazol-3-amine) and poly(9,9-dihexyl-9H-fluoren-2-amine) are prepared by facile chemical polymerization. Then they are characterized with NMR (Nuclear Magnetic Resonance), GPC (Gel Permeation Chromatography), XRD (X-Ray Diffraction), FT-IR (Fourier Transform Infrared spectroscopy), FL (Fluorescence spectrometry) and UV-vis (Ultraviolet-visible spectroscopy) characterizations and further applied to the fluorescence detection of different acids and amines. Moreover, the obtained poly(9-methyl-9H-carbazol-3-amine) displays excellent fluorescence properties in the detection for both acids and amines. Besides, this poly(9-methyl-9H-carbazol-3-amine) can not only be used for fluorescence detection in solution, but also can be prepared into solid state and applied in the gas phase fluorescence detection. This work has greatly expanded the scope of application to these polyaniline derivatives materials, opening a new path for the researches on multi-functional chemosensor.

Excitation wavelength dependence of the fluorescence lifetime of anisole.

Photo-physical models that describe the pressure- and temperature-dependent fluorescence quantum yield of organic fluorescence tracers rely on an accurate prediction of the initial excited-state population, collision-dependent relaxation processes, and state-dependent relaxation processes. In case the initial excited-state population distribution reached after the laser excitation equals on average the thermal distribution, the fluorescence quantum yield becomes pressure independent. This initial distribution critically depends on the temperature-dependent ground-state population before excitation as well as the excitation wavelength. The ability to predict this behavior is a critical check for the validity of the existing photophysical models. The dependence of the effective fluorescence lifetime of anisole on the excitation wavelength (256-270 nm) was investigated at temperatures between 325 and 525 K for pressures between 1 and 4 bar. For each temperature, a unique excitation wavelength was found where the fluorescence lifetime is pressure-independent. The comparison of the experimental results with the predictions based on the established photophysical step-ladder models revealed a systematic underestimation of the required excitation photon energies for direct excitation into the thermalized level. An improved modeling approach based on quantum chemistry calculations for implementing simulated excitation spectra and state-dependent transition probabilities overcomes these limitations. Our results show for the example of anisole that the fluorescence step-ladder models that exist for aromatic fluorescence tracers must be modified to correctly predict the effect of the excitation wavelength.

Laser-induced- and dispersed-fluorescence studies of rhodamine 590 and 640 ions formed by electrospray ionization: observation of fluorescence from highly-excited vibrational levels of S1 states.

The gas-phase fluorescence excitation and emission properties of rhodamine 590 (R590) and rhodamine 640 (R640) cations were studied using a novel instrument that allows extension of optical excitation to the UV region. Ions were generated by an electrospray ion source, and desolvated ions were accumulated in a room-temperature quadrupole ion-trap. In addition to the well-known S1-S0 transition observed for both dyes, two ultraviolet (UV) bands (that have not been reported previously) were observed for R590. When compared with transitions observed for the dyes in methanol solutions, the S1-S0 transitions in the gas phase are shifted to significantly higher energies. These shifts suggest that the S1 states for both dyes are more stabilized by favorable solvent interactions than the S0 states. The UV bands observed for R590 were assigned to the transitions involving the higher-energy S3 and Sn states. The DF spectra obtained upon UV excitation are similar to those recorded for S1 excitation; however, the emission after UV excitation is shifted slightly to the red region. These observations are consistent with the emission from high-vibrational levels of the S1 state formed upon fast internal conversion from S3 or Sn. In vacuo, the absence of collisions extends the lifetimes of vibrationally-excited S1 states, allowing fluorescence from these states to be observed. A similar DF spectrum was also observed for the UV excitation of R640, suggesting that fluorescence from vibrationally hot S1 states is a general phenomenon for ions in the gas phase.

Development of laser-induced fluorescence to quantify in-cylinder fuel wall films

Laser-induced fluorescence of a fuel tracer is a very sensitive technique to image in-cylinder liquid fuel films, but quantification of the measured film thickness has proven difficult so far. This article describes improvements in the quantification procedure and presents an example application in a motored, optically accessible spark-ignition engine with direct injection. We designed a calibration tool that could be pressurized and heated, allowing investigation of the laser-induced fluorescence intensities at temperatures exceeding the liquid’s standard-pressure boiling point. The fluorescence intensity of liquid toluene and 3-Pentanone dissolved in isooctane upon excitation with a pulsed laser at 266 nm was investigated as a function of temperature and pressure. Consistent with the literature results on gas-phase laser-induced fluorescence, the signal from toluene was much stronger than from 3-Pentanone, about two orders of magnitude for films thinner than 50 μm. Laser-induced fluorescence from both tracers decreased with increasing temperature but that of toluene significantly more. The response to pressure was less pronounced. For imaging across a large field of view, the spatial non-uniformity of laser excitation and detection efficiency was taken into account using a solid fluorescing substrate, an inexpensive Schott-glass WG280 filter. Isooctane with 0.5 vol.% toluene was used for application in the motored engine, imaging the liquid film on the piston-top window after direct injection from a central multi-hole injector. Air as a bulk gas was found to be advantageous over nitrogen in that gas-phase fluorescence was quenched by oxygen. The imaged film distributions and thicknesses and the derived total fuel film mass were physically plausible. Consistent with the recent literature results from a constant pressure vessel, increasing injection pressure from 50 to 100 bar did not decrease wall wetting but further increase to 200 bar did.

A cylindrical quadrupole ion trap in combination with an electrospray ion source for gas-phase luminescence and absorption spectroscopy.

A relatively simple setup for collection and detection of light emitted from isolated photo-excited molecular ions has been constructed. It benefits from a high collection efficiency of photons, which is accomplished by using a cylindrical ion trap where one end-cap electrode is a mesh grid combined with an aspheric condenser lens. The geometry permits nearly 10% of the emitted light to be collected and, after transmission losses, approximately 5% to be delivered to the entrance of a grating spectrometer equipped with a detector array. The high collection efficiency enables the use of pulsed tunable lasers with low repetition rates (e.g., 20 Hz) instead of continuous wave (cw) lasers or very high repetition rate (e.g., MHz) lasers that are typically used as light sources for gas-phase fluorescence experiments on molecular ions. A hole has been drilled in the cylinder electrode so that a light pulse can interact with the ion cloud in the center of the trap. Simulations indicate that these modifications to the trap do not significantly affect the storage capability and the overall shape of the ion cloud. The overlap between the ion cloud and the laser light is basically 100%, and experimentally >50% of negatively charged chromophore ions are routinely photodepleted. The performance of the setup is illustrated based on fluorescence spectra of several laser dyes, and the quality of these spectra is comparable to those reported by other groups. Finally, by replacing the optical system with a channeltron detector, we demonstrate that the setup can also be used for gas-phase action spectroscopy where either depletion or fragmentation is monitored to provide an indirect measurement on the absorption spectrum of the ion.

Ion Mobility Spectrometry Coupled to Laser-Induced Fluorescence

We report on interfacing a differential mobility analyzer (DMA) with laser-induced fluorescence (LIF) to simultaneously retrieve two-dimensional information on the electrical mobility and fluorescence spectroscopy of gas-phase ions. The fact that the separation of ions within DMA takes place in space rather than in time allows for the continuous selection of ion beams within a narrow range of mobilities that are further analyzed by LIF. Combination of DMA with LIF is simple and robust. It allows one to detect fluorescence from specified ions, including clusters, which would not survive in a mass spectrometer. Complex mixtures of fluorescent compounds can be separated by the DMA and studied by LIF. LIF is a sensitive technique and useful in the study of molecular interactions. DMA with LIF detection can be used for studies of gas-phase fluorescence of small molecules such as different dyes and their conjugates. This unique instrument combination may also provide a powerful platform for probing fluorescent proteins in the gas phase, which is of great fundamental interest for better understanding of their physical and chemical properties. In the present work, we have studied the gas-phase laser-induced fluorescence of mobility-selected rhodamine 6G ions.

Fluorescence lifetimes of rhodamine dyes in vacuo

Abstract Laser-induced fluorescence lifetimes are reported for several mass-selected gas-phase rhodamine cations, which were generated using electrospray ionization. A recently developed instrument consisting of a quadruple ion trap mass spectrometer coupled to a tunable laser excitation source is used for ion storage and irradiation, respectively. Time-resolved fluorescence measurements are carried out using time-correlated single photon counting (TCSPC), with a Single Photon Avalanche Diode (SPAD) for efficient detection of the light. The time-resolved data are well fit using single exponential decays. Measured lifetimes are 5.65 ± 0.04 ns for gaseous cationic rhodamine 575 (also known as rhodamine 19), 5.83 ± 0.06 ns for rhodamine 590, 5.87 ± 0.05 ns for rhodamine 6G and 6.92 ± 0.06 ns for rhodamine 640 (also known as rhodamine 101). These values are the first reports of the lifetimes of gaseous cationic rhodamine 575, 590 and 6G, while the uncertainty for the reported lifetime of 640 is an order of magnitude smaller than the previously reported value. The reported gas-phase fluorescence lifetimes for these rhodamines are all longer than their measured lifetimes in solution. Photo-physical processes leading to the increase in the fluorescence lifetimes of molecules in the gas phase compared to their solution behavior are discussed. Knowledge of the intrinsic lifetimes of rhodamines, as provided in this work, will aid the development of fluorescence resonance energy transfer as a structural probe of gaseous ions. It will also form a basis from which to better understand how interactions with solvent influence the properties of fluorescent dyes.

The fluorescence properties of cationic rhodamine B in the gas phase

Studying the photophysical properties of molecules in the gasphase can be advantageous, because it reduces the complexity of the system examined by eliminating effects due to theinteractions between the molecule of interest and other species present in the local environment, including those with the solvent itself. Here, we report on theintrinsic properties of gaseous protonated rhodamine B (RBH +), a well-known xanthene-based dye. Protonated rhodamine B was transferred into the gas phase using electrospray ionization (ESI) and isolated in a quadrupole ion trap (QIT) mass spectrometer, which has been modified to enable laser-induced fluorescence spectroscopy of trapped ions. The gas-phase fluorescence excitation and emission spectra of RBH +show maxima ( λ ex (max) = 531 nm and λ em (max) = 542 nm, respectively)that lie at higher energy than those of RBH + in solution. The fluorescence lifetime of gaseous RBH + is 5.97 ± 0.12 ns, which is significantly longer than that of solution-phase rhodamine B. Gaseous rhodamine B is signficiantly brighter than monoethylamino rhodamines such as gaseous rhodamine 6G. Knowledge of the intrinsic photophysical properties of chromophores, such as those presented here for rhodamine B, will enable a better understanding of how the local environment of the chromophore modulates its properties.