Low-temperature Fluorescence Research Articles

De-etiolation is Almost Colour Blind: the Study of Photosynthesis Awakening Under Blue and Red Light.

The synthesis and assembly of functioning photosynthetic complexes in chloroplasts developing from etioplasts during the de-etiolation of angiosperm seedlings are imperative for the plant's autotrophic lifestyle. This study compared de-etiolation process under monochromatic red or blue light of equal photon flux density during a 24-hour illumination period of etiolated Arabidopsis seedlings. The aim was to elucidate the impact of these light wavelength on the etioplast-to-chloroplast transformation and the initiation of light-dependent photosynthetic reactions. Both treatments lead to the formation of functional young chloroplasts; however, the etioplast-to-chloroplast transition and the assembly of photosynthetic complexes occurred unevenly, with individual steps tuned by red or blue light. Ultrastructural analysis suggested faster prolamellar bodies disassembly under blue light, while low temperature fluorescence studies indicated a slower transformation of protochlorophyllide to chlorophyllide, and chlorophyll a, under these conditions. Red light further promoted the synthesis of chlorophyll b and LHCII antenna proteins. However, the efficiency of antennae in dissipating excess absorbed energy was higher for seedlings de-etiolated under blue light; the maximum quantum yield of the photosystem II reached 0.81 after 24-hour de-etiolation, equivalent to mature plants. Blue light seemed to enhance the development of well-functioning photosystems (I and II) and antennae. These findings are important for gaining a deeper understanding of photoreceptor regulation of de-etiolation and for utilizing selected light regimes to improve crop yield.

The effect of magnesium impurities on the optical and spectral characteristics of calcium orthovanadate single crystals doped with chromium and manganese

The effect of magnesium co-doping on growth conditions, crystal composition, optical and spectral properties of manganese or chromium-doped Ca3(VO4)2 (CVO) crystal was studied. The single crystals were grown by the Czochralski method. Effective segregation coefficients of manganese and chromium in the CVO matrix with magnesium co-doping were calculated. A redistribution of the absorption lines intensities was observed in the absorption spectra of CVO:Mn:Mg and CVO:Cr:Mg. Low (15 K) temperature fluorescence measurements confirmed that co-doping CVO crystal with Mg ions leads to the redistribution of Mn ions concentration in different valence states, but does not cause changes in their spectral properties. In the presence of magnesium, additional annealing of the materials resulted in a slower reduction of the valence state of doping ions and a noticeable change in crystal coloration. All investigated impurities have practically no effect on the specific ferroelectric domain structure of CVO.

Bromine Atoms Decorated Pyene-based Covalent Organic Frameworks for Accelerated Photocatalytic H2 Production.

Designing materials with low exciton binding energy is an efficient way of improving the hydrogen production performance of COFs（Covalent Organic Frameworks. Here, it is demonstrated that the strategy of decorating bromine atoms on Pyene-based COFs can achieve elevated photocatalytic H2 evolution rates (HER = 13.61mmol g-1h-1). Low-temperature fluorescence and time-resolved fluorescence spectroscopy (TRPL) indicate that the introduction of bromine atoms can significantly suppress charge recombination. DFT (Density Functional Theory) calculation clarified that the C atoms adjacent to Br are the active sites with a reduced energy barrier in the process of formatting H intermediate species (H*). The modification strategy of Br atoms in COF furnishes a new medium for exploiting exquisite photocatalysts.

Phycobilisome protein ApcG interacts with PSII and regulates energy transfer in Synechocystis.

Photosynthetic organisms harvest light using pigment-protein complexes. In cyanobacteria, these are water-soluble antennae known as phycobilisomes (PBSs). The light absorbed by PBS is transferred to the photosystems in the thylakoid membrane to drive photosynthesis. The energy transfer between these complexes implies that protein-protein interactions allow the association of PBS with the photosystems. However, the specific proteins involved in the interaction of PBS with the photosystems are not fully characterized. Here, we show in Synechocystis sp. PCC 6803 that the recently discovered PBS linker protein ApcG (sll1873) interacts specifically with PSII through its N-terminal region. Growth of cyanobacteria is impaired in apcG deletion strains under light-limiting conditions. Furthermore, complementation of these strains using a phospho-mimicking version of ApcG causes reduced growth under normal growth conditions. Interestingly, the interaction of ApcG with PSII is affected when a phospho-mimicking version of ApcG is used, targeting the positively charged residues interacting with the thylakoid membrane, suggesting a regulatory role mediated by phosphorylation of ApcG. Low-temperature fluorescence measurements showed decreased PSI fluorescence in apcG deletion and complementation strains. The PSI fluorescence was the lowest in the phospho-mimicking complementation strain, while the pull-down experiment showed no interaction of ApcG with PSI under any tested condition. Our results highlight the importance of ApcG for selectively directing energy harvested by the PBS and imply that the phosphorylation status of ApcG plays a role in regulating energy transfer from PSII to PSI.

Water-Deficit Stress in the Epiphytic Elkhorn Fern: Insight into Photosynthetic Response.

Progressive climate changes cause disturbance of water relations in tropical rainforests, where epiphytic ferns are an important element of biodiversity. In these plants, the efficiency of photosynthesis is closely related to the efficiency of water transport. In addition, due to the lack of contact with the soil, epiphytes are extremely susceptible to water-deficit stress. The aim of this experiment was to determine the response of the photosynthetic apparatus of Platycerium bifurcatum to a 6-week water deficit. The hydration and pigment composition of leaves were determined using reflectance spectroscopy and epifluorescence microscopy. Chlorophyll a fluorescence kinetics parameters, fluorescence induction curves (OJIP), low-temperature fluorescence curves at 77 K and proline concentration were analyzed at seven time points. After a decrease in leaf hydration by 10-15%, there were disturbances in the oxidation-reduction balance, especially in the initial photochemical reactions, a rapid decrease in plant vitality (PI) and significant fluctuations in chlorophyll a fluorescence parameters. The relative size of PSI antenna structures compared to PSII decreased in the following weeks of water deficit. Changes in photochemical reactions were accompanied by a decrease in gross photosynthesis and an increase in proline concentration. Changes in the functioning of photosynthesis light phase and the pigment composition of leaves are related to the resistance of elkhorn fern to long-term water deficit.

Triplet-Induced Singlet Oxygen Photobleaches Near-Infrared Dye-Sensitized Upconversion Nanosystems.

The rapid photobleaching of near-infrared (NIR) dye-sensitized upconversion nanosystems is one of the crucial problems that has blocked their technological applications. Uncovering the photophysical and photochemical pathways of NIR dyes would help to elucidate the photobleaching mechanism and thereby improve the photostability of the system. Here we investigate the triplet dynamics of NIR dyes and their interaction with triplet oxygen in the typically investigated IR806-sensitized upconversion nanoparticle (UCNP) nanosystem. Low-temperature fluorescence at 77 K provides direct proof of the generation of singlet oxygen (1O2) under 808 nm laser irradiation. Mass spectrometry indicates that all three double bonds in the structure of IR806 can be broken in the photochemical process. Coupling IR806 to the surface of UCNPs can accelerate its triplet dynamics, thus producing more 1O2 to photocleave IR806. Importantly, we find that the addition of β-carotene can scavenge the generated 1O2, thereby providing a simple method to effectively inhibit photobleaching.

Activating ultralow upconversion nanothermometry in neodymium sublattice for heart tissue imaging rapid-responsive

The fields of biosensitivity and biological imaging have received a lot of attention from rare earth-doped upconversion nanoparticles (UCNPs). However, owing to the relatively large energy difference of rare earth ions, biological sensitivity based on UCNPs is restricted to detect at low temperature. Here, we design core-shell-shell NaErF4:Yb@Nd2O3@SiO2 UCNPs as a dual-mode bioprobe that produces blue, green, and red multi-color upconversion emissions at extremely low temperatures between 100 K and 280 K. Based on the thermally coupled energy levels (TCELs) of Er3+ (2H11/2 and 4S3/2) and Nd3+ (4F5/2 and 4F3/2) at 100 K, the greatest relative sensitivity (SR) approaches 12.7% K−1. NaErF4:Yb@Nd2O3@SiO2 injection is used to achieve blue upconversion emission imaging of frozen heart tissue, showing that this UCNP can serve as a low-temperature sensitive biological fluorescence.

Long- and short-term acclimation of the photosynthetic apparatus to salinity in Chlamydomonas reinhardtii. The role of Stt7 protein kinase.

Salt stress triggers an Stt7-mediated LHCII-phosphorylation signaling mechanism similar to light-induced state transitions. However, phosphorylated LHCII, after detaching from PSII, does not attach to PSI but self-aggregates instead. Salt is a major stress factor in the growth of algae and plants. Here, our study mainly focuses on the organization of the photosynthetic apparatus to the long-term responses of Chlamydomonas reinhardtii to elevated NaCl concentrations. We analyzed the physiological effects of salt treatment at a cellular, membrane, and protein level by microscopy, protein profile analyses, transcripts, circular dichroism spectroscopy, chlorophyll fluorescence transients, and steady-state and time-resolved fluorescence spectroscopy. We have ascertained that cells that were grown in high-salinity medium form palmelloids sphere-shaped colonies, where daughter cells with curtailed flagella are enclosed within the mother cell walls. Palmelloid formation depends on the presence of a cell wall, as it was not observed in a cell-wall-less mutant CC-503. Using the stt7 mutant cells, we show Stt7 kinase-dependent phosphorylation of light-harvesting complex II (LHCII) in both short- and long-term treatments of various NaCl concentrations-demonstrating NaCl-induced state transitions that are similar to light-induced state transitions. The grana thylakoids were less appressed (with higher repeat distances), and cells grown in 150 mM NaCl showed disordered structures that formed diffuse boundaries with the flanking stroma lamellae. PSII core proteins were more prone to damage than PSI. At high salt concentrations (100-150 mM), LHCII aggregates accumulated in the thylakoid membranes. Low-temperature and time-resolved fluorescence spectroscopy indicated that the stt7 mutant was more sensitive to salt stress, suggesting that LHCII phosphorylation has a role in the acclimation and protection of the photosynthetic apparatus.

Preparation and curing properties of waterborne epoxy emulsified asphalt/DMP-30 composites

The objective of this paper is to investigate the effect of DMP-30 on the curing rate and mechanical properties of waterborne epoxy emulsified asphalt (WEREA). WEREA/DMP-30 composites were prepared and the high temperature rheological properties, low temperature rheological properties, adhesion properties, tensile properties and fluorescence microscopy images of DMP-30 modified WEREA were investigated. Results showed that DMP-30 can effectively reduce the curing time of WEREA. There are positive and negative effects of DMP-30 on the mechanical properties of WEREA.The 1% and 2% DMP-30 dosing showed an enhancement of the high temperature performance and tensile strength, while the 3% DMP-30 dosing showed a weakening effect. In addition, the ductility and flexibility of WEREA are improved with the addition of DMP-30. The above results provide a reference for material optimization design of waterborne epoxy resin modified emulsified asphalt.

Fluorescence Lifetimes of NIR-Emitting Molecules with Excited-State Intramolecular Proton Transfer

Molecular probes based on the excited-state intramolecular proton-transfer (ESIPT) mechanism have emerged to be attractive candidates for various applications. Although the steady-state fluorescence mechanisms of these ESIPT-based probes have been reported extensively, less information is available about the fluorescence lifetime characteristics of newly developed NIR-emitting dyes. In this study, four NIR-emitting ESIPT dyes with different cyanine terminal groups were investigated to evaluate their fluorescence lifetime characteristics in a polar aprotic solvent such as CH2Cl2. By using the time-correlated single-photon counting (TCSPC) method, these ESIPT-based dyes revealed a two-component exponential decay (τ1 and τ2) in about 2-4 nanoseconds (ns). These two components could be related to the excited keto tautomers. With the aid of model compounds (5 and 6) and low-temperature fluorescence spectroscopy (at -189 ℃), this study identified the intramolecular charge transfer (ICT) as one of the major factors that influenced the τ values. The results of this study also revealed that both fluorescence lifetimes and fractional contributions of each component were significantly affected by the probe structures.

The location of the low-energy states in Lhca1 favors excitation energy transfer to the core in the plant PSI-LHCI supercomplex.

Lhca1 is one of the four pigment-protein complexes composing the outer antenna of plant Photosystem I-light-havesting I supercomplex (PSI-LHCI). It forms a functional dimer with Lhca4 but, differently from this complex, it does not contain 'red-forms,' i.e., pigments absorbing above 700nm. Interestingly, the recent PSI-LHCI structures suggest that Lhca1 is the main point of delivering the energy harvested by the antenna to the core. To identify the excitation energy pathways in Lhca1, we developed a structure-based exciton model based on the simultaneous fit of the low-temperature absorption, linear dichroism, and fluorescence spectra of wild-type Lhca1 and two mutants, lacking chlorophylls contributing to the long-wavelength region of the absorption. The model enables us to define the locations of the lowest energy pigments in Lhca1 and estimate pathways and timescales of energy transfer within the complex and to the PSI core. We found that Lhca1 has a particular energy landscape with an unusual (compared to Lhca4, LHCII, and CP29) configuration of the low-energy states. Remarkably, these states are located near the core, facilitating direct energy transfer to it. Moreover, the low-energy states of Lhca1 are also coupled to the red-most state (red forms) of the neighboring Lhca4 antenna, providing a pathway for effective excitation energy transfer from Lhca4 to the core.

Zam Is a Redox-Regulated Member of the RNB-Family Required for Optimal Photosynthesis in Cyanobacteria.

The zam gene mediating resistance to acetazolamide in cyanobacteria was discovered thirty years ago during a drug tolerance screen. We use phylogenetics to show that Zam proteins are distributed across cyanobacteria and that they form their own unique clade of the ribonuclease II/R (RNB) family. Despite being RNB family members, multiple sequence alignments reveal that Zam proteins lack conservation and exhibit extreme degeneracy in the canonical active site—raising questions about their cellular function(s). Several known phenotypes arise from the deletion of zam, including drug resistance, slower growth, and altered pigmentation. Using room-temperature and low-temperature fluorescence and absorption spectroscopy, we show that deletion of zam results in decreased phycocyanin synthesis rates, altered PSI:PSII ratios, and an increase in coupling between the phycobilisome and PSII. Conserved cysteines within Zam are identified and assayed for function using in vitro and in vivo methods. We show that these cysteines are essential for Zam function, with mutation of either residue to serine causing phenotypes identical to the deletion of Zam. Redox regulation of Zam activity based on the reversible oxidation-reduction of a disulfide bond involving these cysteine residues could provide a mechanism to integrate the ‘central dogma’ with photosynthesis in cyanobacteria.

Assembly of LHCA5 into PSI blue shifts the far-red fluorescence emission in higher plants

In higher plants, the PSI core complex is associated with light-harvesting complex I (LHCI), forming the PSI-LHCI super-complex. In vascular plants, four major antenna proteins (LHCA1-4) are assembled in the order of LHCA1, LHCA4, LHCA2, and LHCA3 into a crescent-shaped LHCI, while LHCA5 and LHCA6 are minor antenna proteins. By contrast, in moss and green algae, LHCA5 or LHCA5-like protein functions as one of the major antenna proteins by residing at the second site of LHCI. In order to learn the effect of binding different LHCA proteins, i.e. LHCA4 or LHCA5, within the PSI-LHCI super-complex on photosynthetic properties of plants, we constructed LHCA5 overexpression plants with a wild type (WT) background and an lhca4 mutant background in Arabidopsis thaliana. The results showed that: (i) there are little difference in phenotype, pigment composition and chlorophyll fluorescence parameters between the transgenic Arabidopsis and their corresponding background materials; (ii) in spite of a small amount of LHCA5, the LHCA5-included PSI-LHCI super-complex can be obtained by extracting samples incubated with anti-FLAG M2 Affinity Gel, in which LHCA5 is found to substitute for LHCA4 as analyzed by immunoblotting analysis; (iii) the replacement of LHCA4 with LHCA5 within PSI-LHCI super-complex leads to a blue shift in low temperature fluorescence emission, suggesting a decrease in far-red absorbance. These results provide new clues for understanding the position and function of LHCA4 and LHCA5 during the evolution of green plants from aquatic to terrestrial lifestyles.

Oxidative Stress- and Autophagy-Inducing Effects of PSI-LHCI from Botryococcus braunii in Breast Cancer Cells.

Botryococcus braunii (B. braunii) is a green microalga primarily found in freshwater, reservoirs, and ponds. Photosynthetic pigments from algae have shown many bioactive molecules with therapeutic potential. Herein, we report the purification, characterization, and anticancer properties of photosystem I light-harvesting complex I (PSI-LHCI) from the green microalga B. braunii UTEX2441. The pigment–protein complex was purified by sucrose density gradient and characterized by its distinctive peaks using absorption, low-temperature (77 K) fluorescence, and circular dichroism (CD) spectroscopic analyses. Protein complexes were resolved by blue native-PAGE and two-dimensional SDS-PAGE. Triple-negative breast cancer MDA-MB-231 cells were incubated with PSI-LHCI for all of our experiments. Cell viability was assessed, revealing a significant reduction in a time- and concentration-dependent manner. We confirmed the internalization of PSI-LHCI within the cytoplasm and nucleus after 12 h of incubation. Cell death mechanism by oxidative stress was confirmed by the production of reactive oxygen species (ROS) and specifically superoxide. Furthermore, we monitored autophagic flux, apoptotic and necrotic features after treatment with PSI-LHCI. Treated MDA-MB-231 cells showed positive autophagy signals in the cytoplasm and nucleus, and necrotic morphology by the permeabilization of the cell membrane. Our findings demonstrated for the first time the cytotoxic properties of B. braunii PSI-LHCI by the induction of ROS and autophagy in breast cancer cells.

The phosphatase/kinase balance affects phytochrome A and its native pools, phyA' and phyA″, in etiolated maize roots: evidence from the induction of phyA' destruction by a protein phosphatase inhibitor sodium fluoride.

Phytochrome A (phyA) comprises two native types, phyA' and phyA″, with distinct spectroscopic, photochemical, and functional properties, differing at the N-terminal extension, probably, by the state of phosphorylation. To find out if and how protein phosphatases (PP) affect the state of the phyA species in planta, we studied the effect of the non-specific phosphatase inhibitor NaF on etiolated maize seedlings with the use of low-temperature fluorescence spectroscopy and photochemistry. In roots, phosphatase inhibition facilitated photoreceptor destruction in its labile phyA' form and shifted the phyA'/phyA″ ratio towards the more stable phyA″. The effect of NaF was not observed in stems. It was similar, though less pronounced, in comparison to the effects of the serine/threonine PP inhibitors, okadaic and cantharidic acids (OA and CA), which likewise facilitate the destruction of phyA' in etiolated maize stems, not, however, in roots (Sineshchekov et al., Photochem. Photobiol 89:83-96, 2013). The phyA'/phyA″ balance thus depends on the kinase/phosphatase equilibrium in the root cells. The relatively low effect of NaF on phyA in roots, together with the lack of the effect of OA and CA in them, may imply that the mechanism controlling the phyA'/phyA″ balance in roots can be different from that in shoots.

Unravelling the ambiguity of the emission pattern of donor–acceptor salicylaldimines

Four donor–acceptor salicylaldimines with benzoheterocyclic substituents at nitrogen imine were prepared and characterised by means of optical spectroscopy. Their luminescent properties were compared with two isoelectronic carbocyclic analogues. In contrast to the typical salicylaldimines, our reported benzoheterocyclic compounds exhibit solely local emissive behaviour in all media whereas for the carbocyclic analogues we expectedly observe dominant ESIPT emission in the majority of instances. Low temperature (5 K - liquid helium) fluorescence measurements revealed the effect of the restriction of the molecular rotation which was manifested by significant increment of the fluorescence intensity. The comprehensive experimental and ab initio study reveal effect of solvent polarity, viscosity and temperature on the complex photophysics of heterocyclic and carbocyclic substituted salicylaldimines with several degrees of freedom for molecular transformations. This work adds considerable insight in emission lifetime data which greatly facilitates understanding of the fluorescence mechanism of the molecules with environmentally dependent restrictions.

Thermal Stress, Aggregation of Chlorophyll-Protein Complexes, and Light-Dependent Recovery of PSII activity in Wheat Seedlings

The dynamics of changes in the photochemical activity of photosystem II (PSII) and low-temperature spectra at 77 K in the first leaves of 11-day winter wheat plants Triticum aestivum L., as well as structural changes in chlorophyll-protein complexes (CPC) of thylakoid membranes during recovery after a short-term (20 min) heating at a temperature of 42°C, were studied. Changes in the Fv/Fm, F735/F695, and F735/F685 ratios indicate inhibition of PSII immediately after heating. Using nondenaturing electrophoresis, it was shown that the light-harvesting Chl a/b complex of PSII does not aggregate immediately after heating but after several hours, after 6 h the desagregation of CPC was observed, which was consistent with an increase in the Fv/Fm ratio upon recovery. The influence of temperature, intensity, and quality of light (white, blue, and red light) on the recovery of PSII activity and low-temperature fluorescence spectra was studied. It was concluded that the recovery is a photo-activated low-energy process, independent of photosynthesis, and the most effective in blue light.

Combined spectroscopic and DFT studies of local defect structures in beta-tricalcium phosphate doped with Nd(III)

We investigated the distribution of the Nd3+ impurity centers over five Ca2+ positions in the β-Ca3(PO4)2 lattice (space group R3c) using low-temperature site-selective laser spectroscopy and fluorescence kinetics measured by double spectral selection in combination with the results of periodic density functional theory (DFT) calculations. Site-selective fluorescence excitation spectra of the Nd3+ ion were measured by fine tuning the laser wavelength in the spectral band of the 4I9/2(1) → 4G5/2(1,2) transitions and by fluorescence detection in the spectral band of the 4F3/2(1) → 4I9/2(1,2) transitions. The kinetics of nonradiatiative energy transfer from the 4F3/2(1) crystal field (CF) level was used to probe the local structure of Nd3+ sites. Theoretical modelling of the replacement of Ca2+ ions by Nd3+ ions with different charge compensation schemes was carried out. In the context of heterovalent substitution, hydrothermal synthesis conditions, and charge-balance mechanisms, two structural models of substitution have been proposed and thoroughly studied: substitution by a single Nd3+ ion accompanied by a trapped hydroxyl group, and substitution with a pair of Nd3+ ions. Numerical modeling of possible substitution scenarios calculated for a large number of combinations of cation-exchange sites in the lattice was carried out. The results obtained were classified according to the parameters of the accommodation of impurities and the energies of defect formation. The most favorable configurations of the distribution of impurity Nd3+ cations in the β-Ca3(PO4)2 lattice were predicted. By combining the “energy transfer probe” analysis and the results of the structural modeling, a relationship was found between the local geometry and the spectral and kinetic properties of luminescence of the Nd3+ optical centers. This allowed us to accurately determine the local structural geometries of eight experimentally detected the Nd3+ optical centers among the various possibilities in the distribution of the dopant ions over five cationic positions in β-Ca3(PO4)2. Six optical centers represent different types of Nd3+ - Nd3+ pairs, the formation of which is energetically favorable for charge compensation according to the 3Ca2+ → 2Nd3+ + ⎕ scheme, when one calcium position remains vacant. Two optical centers are single centers of the Nd3+ ion in which the charge is compensated by negatively charged hydroxyl ions intercalated during hydrothermal synthesis according to the scheme Ca2+→ Nd3+ + OH−. It was assumed that these pairs are located in several structural cavities free of PO4 tetrahedra.

Electronic spectra and structure of 1- and 2-fluoronaphthalene impurity centers in crystalline naphthalene

The results of experimental investigations on the low-temperature fluorescence and absorption spectra of the 1- and 2-fluoronaphthalene impurities in the crystalline naphthalene are presented and analyzed. When the 1- and 2-fluoronaphthalene concentrations are less than 1%, the absorption and fluorescence spectra are shown to be composed of two identical band series, induced by the formation of two different centers, consisting of the single impurity molecules. The impurity concentration increase of over 1% results in the appearance of the additional structure in the polarized absorption spectra near the 0-0 bands of the single centers in a form of narrow bands with similar intensity and different polarization. This structure is associated with a formation of impurity paired centers, consisting of the two resonantly interacting molecules of either 1-fluoronaphthalene, or 2-fluoronaphthalene. Besides, in the case of the 2-fluoronaphthalene, the absorption spectra contain intense peaks, located ∼ 45 cm−1 higher in frequency, relative to the 0−0 transition bands for each single-center, and consisted of complexes of narrow lines. These spectral peaks are also explained by a formation of impurity paired centers, but here interactions between the molecules are not resonant. For the pure naphthalene crystal factors, causing the observation of additional oppositely polarized weak bands in the absorption spectra at ∼ 10 cm−1 near the Davydov doublet components, are described.

Low-temperature and low-pressure effective fluorescence lifetimes and spectra of gaseous anisole and toluene

Fluorescence spectra and lifetimes of anisole and toluene vapor in nitrogen have been measured at conditions below ambient (257–293 K and 100–2000 mbar) upon excitation with 266-nm laser light to expand the applicable range of anisole and toluene laser-induced fluorescence (LIF) for conditions below room temperature that occur in expanding flows and cases with strong evaporative cooling. Anisole fluorescence spectra broaden with decreasing pressure while fluorescence lifetimes decrease simultaneously. This is consistent with a more pronounced effect of internal vibrational redistribution on the overall fluorescence signal and can be explained by significantly reduced collision rates. In the case of toluene, the transition from photo-induced heating to photo-induced cooling was observed for the first time for 266 nm. The data confirm predictions of earlier work and is particularly important for the advancement of the available photo-physical (step-ladder) models: since those transitions mark points where the molecules are already thermalized after excitation (i.e., no vibrational relaxation occurs during deactivation), they are important support points for fitting empirical parameters and allow analytical determination of the ground state energy transferred to the excited state. The data enable temperature and/or pressure sensing, e.g., in accelerating cold flows using laser-induced fluorescence of both tracers.