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The ability to detect and inactivate spore-forming bacteria is of significance within, for example, industrial, healthcare, and defense sectors. Not only are stringent protocols necessary for the inactivation of spores but robust procedures are also required to detect viable spores after an inactivation assay to evaluate the procedure’s success. UV radiation is a standard procedure to inactivate spores. However, there is limited understanding regarding its impact on spores’ spectral and morphological characteristics. A further insight into these UV-induced changes can significantly improve the design of spore decontamination procedures and verification assays. This work investigates the spectral and morphological changes to Bacillus thuringiensis spores after UV exposure. Using absorbance and fluorescence spectroscopy, we observe an exponential decay in the spectral intensity of amino acids and protein structures, as well as a logistic increase in dimerized DPA with increased UV exposure on bulk spore suspensions. Additionally, using micro-Raman spectroscopy, we observe DPA release and protein degradation with increased UV exposure. More specifically, the protein backbone’s 1600–1700 cm^–1 amide I band decays slower than other amino acid-based structures. Last, using electron microscopy and light scattering measurements, we observe shriveling of the spore bodies with increased UV radiation, alongside the leaking of core content and disruption of proteinaceous coat and exosporium layers. Overall, this work utilized spectroscopy and electron microscopy techniques to gain new understanding of UV-induced spore inactivation relating to spore degradation and CaDPA release. The study also identified spectroscopic indicators that can be used to determine spore viability after inactivation. These findings have practical applications in the development of new spore decontamination and inactivation validation methods.

Optical-optical double resonance (OODR) spectroscopy is a powerful tool for the experimental assignment of highly-excited molecular states, which in turn is needed for verification of the accuracy of theoretical predictions of high-temperature spectra observed in exoplanets and in combustion environments. Previous implementations of OODR used either continuous wave (cw) lasers, which limit the number of transitions that can be detected, or pulsed lasers, which limit the spectral resolution. Recently, we demonstrated OODR with a cw pump and a frequency comb probe and applied it to the detection and assignment of methane transitions in the 3ν 3 ← ν 3 range with sub-Doppler resolution over 200 cm -1 of bandwidth [1]. The pump [see Fig. 1(a) ] was a 1 W 3.3m idler of a cw optical parametric oscillator (cw-OPO), stabilized to the Lamb dip in a selected CH 4 transition in the ν 3 band using a signal from a reference cell. The probe was an amplified fully-stabilized Er:fiber comb ( f rep = 250 MHz), whose center wavelength was shifted to 1.67m using a soliton self-frequency shift fiber (SSSF). The sample of pure CH 4 was contained in an 80-cm-long single-pass cell cooled by liquid nitrogen. The probe spectra were detected using a Fourier transform spectrometer (FTS) with comb-mode-limited resolution [2] , and the final interleaved spectra had 2 MHz sampling point spacing. Figure 1(b) shows the 3ν 3 ← ν 3 R(1) transition at 6046.36008(5) cm -1 , detected with the pump on the ν 3 R(0) line. We measured, fit and assigned 36 probe transitions with the pump tuned to 9 different transitions. Figure 1(d) shows a comparison of the probe transition wavenumbers to predictions from the TheoReTS database [3] , demonstrating agreement within 1 cm -1.

Optical-optical double-resonance spectroscopy with a continuous wave pump and frequency comb probe allows measurement of sub-Doppler transitions to highly excited molecular states over a wide spectral range with high frequency accuracy. We report on assessment and characterization of sub-Doppler double-resonance transitions in methane measured using a 3.3-μm continuous wave optical parametric oscillator as a pump and a 1.67-μm frequency comb as a probe. The comb spectra were recorded using a Fourier transform spectrometer with comb-mode-limited resolution. With the pump tuned to nine different transitions in the ν3 fundamental band, we detected 36 ladder-type transitions to the 3ν3 overtone band region, and 18 V-type transitions to the 2ν3 overtone band. We describe in detail the experimental approach and the pump stabilization scheme, which currently limits the frequency accuracy of the measurement. We present the data analysis procedure used to extract the frequencies and intensities of the probe transitions for parallel and perpendicular relative pump-probe polarization. We compare the center frequencies and relative intensities of the ladder-type transitions to theoretical predictions from the TheoReTS and ExoMol line lists, demonstrating good agreement with TheoReTS.

We report the first measurement of sub-Doppler molecular response using a frequency comb by employing the comb as a probe in optical-optical double-resonance spectroscopy. We use a 3.3 μm continuous wave pump and a 1.67 μm comb probe to detect sub-Doppler transitions to the 2ν3 and 3ν3 bands of methane with ∼1.7 MHz center frequency accuracy. These measurements provide the first verification of the accuracy of theoretical predictions from highly vibrationally excited states, needed to model the high-temperature spectra of exoplanets. Transition frequencies to the 3ν3 band show good agreement with the TheoReTS line list.

We use a 3.3 µm high-power continuous wave pump and a 1.67 µm comb probe to detect transitions in the 3ν3 ← ν3 range of methane with sub-Doppler resolution over 6 THz of bandwidth. We achieve high absorption sensitivity for the comb probe using an enhancement cavity and a Fourier transform spectrometer with auto-balanced detection.

We use a 3.3 µm continuous wave optical parametric oscillator as a pump and a 1.67 µm frequency comb as a probe to record 36 sub-Doppler double-resonance transitions in the 3v3 band of methane (including 26 previously unreported) with ~1.5 MHz center frequency accuracy.

We use a 3.3 μm continuous wave optical parametric oscillator as a pump and a 1.67 μm frequency comb as a probe to record 36 sub-Doppler double-resonance transitions in the 3v3 band of methane (including 26 previously unreported) with ∼1.5 MHz center frequency accuracy.

Accurate parameters of molecular transitions are needed for data analysis in many applications, ranging from atmospheric research to astrophysics and determination of fundamental constants. Optical frequency comb Fourier transform spectroscopy (OFC-FTS) is particularly well-suited for high-precision measurements of broadband molecular spectra. From these spectra, the parameters of individual transitions - all measured simultaneously under the same experimental conditions - can be determined. We use a mechanical OFC-FTS spectrometer with sub-nominal resolution [1, 2] to perform precise broadband measurements of entire molecular bands of CO2 using either direct absorption spectroscopy or cavity-enhanced complex refractive index spectroscopy (CE-CRIS) [3] and we extract line parameters for line shapes beyond the Voigt profile.

An absorption spectrum of (H2O)-O-16 at 1950 K is recorded in a premixed methane/air flat flame using a cavity-enhanced optical frequency comb-based Fourier transform spectrometer. 2417 absorption lines are identified in the 6250-6670 cm(-1) region with an accuracy of about 0.01 cm(-1). Absolute line intensities are retrieved using temperature and concentration values obtained by tunable diode laser absorption spectroscopy. Line assignments are made using a combination of empirically known energy levels and predictions from the new POKAZATEL variational line list. 2030 of the observed lines are assigned to 2937 transitions, once blends are taken into account. 126 new energy levels of (H2O)-O-16 are identified. The assigned transitions belong to 136 bands and span rotational states up to J = 27.

We present broadband cavity-enhanced complex refractive index spectroscopy (CE-CRIS), a technique for calibration-free determination of the complex refractive index of entire molecular bands via direct measurement of transmission modes of a Fabry-Perot cavity filled with the sample. The measurement of the cavity transmission spectrum is done using an optical frequency comb and a mechanical Fourier transform spectrometer with sub-nominal resolution. Molecular absorption and dispersion spectra (corresponding to the imaginary and real parts of the refractive index) are obtained from the cavity mode broadening and shift retrieved from fits of Lorentzian profiles to the individual cavity modes. This method is calibration-free because the mode broadening and shift are independent of the cavity parameters such as the length and mirror reflectivity. In this first demonstration of broadband CE-CRIS we measure simultaneously the absorption and dispersion spectra of three combination bands of CO2 in the range between 1525 nm and 1620 nm and achieve good agreement with theoretical models. This opens up for precision spectroscopy of the complex refractive index of several molecular bands simultaneously.