Changes In Fluorescence Intensity Research Articles

Bioimaging research of a turn-on homodimer fluorescent probes for pH and viscosity: A new turn-on strategy

A turn-on homodimer fluorescent probe (BL) that responded to pH and viscosity was facilely synthesized based on homo-FRET. The probe exhibited strong fluorescence intensity and color changes in alkaline pH. In addition, the probe's fluorescence had altered with changes in viscosity. What's more, Mitochondrial localization and pH bioimaging in E. coli cell experiments confirmed that BL was a novel multifunctional probe that could be applied to mitochondrial-targeted fluorescence imaging under normal conditions and real-time fluorescence imaging of E. coli under alkaline conditions.

Eu3+-functionalized covalent organic framework for ratiometric fluorescence detection and adsorption of tetracycline and information steganography.

Functional materials with organic/inorganic composites as the main matrix and rare earth ion complexes as the guest have shown a very broad application prospect for antibiotic sensors. However, Eu3+-complex often relies on a single fluorescence response signal, which is susceptible to changes in the detection environment and cannot simultaneously detect and remove tetracycline (TC). Herein, green fluorescent covalent two-dimensionalorganic framework (COF-TD) is synthesized, followed by modification of Eu3+ to synthesize COF-TD@Eu3+. In the ratiometric sensor, Eu3+ serves as the recognition site and specific response probe for TC, while COF-TD is the fluorescence reference and carrier for Eu3+. Due to the antenna effect, TC enhances the red fluorescence of Eu3+, while the green fluorescence of COF-TD remainsalmost stable. Based on the change of fluorescence intensity and fluorescence color from green to red, the efficient ratiometric sensing can be finished in 1min. The developed method shows high sensitivitywith a detection limit of 0.3μM and highselectivity to TC which makes the methodapplicableto detect TC in traditional Chinese medicine preparations. In addition, due to the high specific surface area of COFs and specific adsorption sites, COF-TD@Eu3+ also shows good performance for TC removal. The findings show that the maximum adsorption capacity is 137.3mgg-1 and the adsorption equilibrium is reached in 30min. Smartphone assisted COF-TD@Eu3+ for both ratiometric fluorescence detection and detecting the absorption ofTC is proposed for the first time. The molecular cryptosteganography that transforms the selective response of COF-TD@Eu3+ to binary strings is anticipated to advance utilization of nanomaterials in logic sensing and information safety.

Probing Temperature‐Dependence of Hydrogen Bonding in Condensed Polymeric Materials with Aggregation‐Induced Emission

AbstractHydrogen bonding is pivotal in chemical and biological systems, yet detecting it in condensed matter, especially polymers, remains challenging. We propose a straightforward method using an aggregation‐induced emission luminogen (AIEgen) probe, demonstrated in a specially designed hydrogen‐bonded polymer with UPy groups. Unlike conventional covalent bonding, our approach involves mixing the AIEgen probe with polymers, inducing a significant change in fluorescence intensity due to the exceptional AIE characteristics. Validation involved comparative analyses with traditional methods, including dynamic mechanical analysis and infrared spectroscopy, across various temperatures. Our AIEgen‐based approach offers a more intuitive assessment of hydrogen bonding dynamics than the complexity of infrared spectroscopy. Importantly, hydrogen‐bonding transition temperatures determined by our AIEgen‐based method are only 6–8 degrees higher than those from dynamic mechanical analyses, confirming accuracy under faster, simpler conditions. This innovative technique not only opens a new avenue for investigating hydrogen bonding dynamics within confined polymer chains but also enhances the precision of transition temperature determination in solid‐state materials. Our study highlights the vital role of hydrogen bonding in functional materials, promising exciting possibilities for further advancements in this field.

Exploring the Role of Neuropeptide PACAP in Cytoskeletal Function Using Spectroscopic Methods.

The behavior and presence of actin-regulating proteins are characteristic of various clinical diseases. Changes in these proteins significantly impact the cytoskeletal and regenerative processes underlying pathological changes. Pituitary adenylate cyclase-activating polypeptide (PACAP), a cytoprotective neuropeptide abundant in the nervous system and endocrine organs, plays a key role in neuron differentiation and migration by influencing actin. This study aims to elucidate the role of PACAP as an actin-regulating polypeptide, its effect on actin filament formation, and the underlying regulatory mechanisms. We examined PACAP27, PACAP38, and PACAP6-38, measuring their binding to actin monomers via fluorescence spectroscopy and steady-state anisotropy. Functional polymerization tests were used to track changes in fluorescent intensity over time. Unlike PACAP27, PACAP38 and PACAP6-38 significantly reduced the fluorescence emission of Alexa488-labeled actin monomers and increased their anisotropy, showing nearly identical dissociation equilibrium constants. PACAP27 showed weak binding to globular actin (G-actin), while PACAP38 and PACAP6-38 exhibited robust interactions. PACAP27 did not affect actin polymerization, but PACAP38 and PACAP6-38 accelerated actin incorporation kinetics. Fluorescence quenching experiments confirmed structural changes upon PACAP binding; however, all studied PACAP fragments exhibited the same effect. Our findings indicate that PACAP38 and PACAP6-38 strongly bind to G-actin and significantly influence actin polymerization. Further studies are needed to fully understand the biological significance of these interactions.

A non-toxic inclusion complexation of loxoprofen with hydroxypropyl-β-cyclodextrin for complete solubility and anti-inflammatory efficacy

In recent days, there has been a significant demand for drugs that are easily soluble, non-toxic, safe, and biocompatible for human use. Recently, active pharmaceutical ingredient (API) complexes with cyclodextrins (CDs) and their derivatives have gained considerable attention. In line with this trend, we have developed an inclusion complex material for loxoprofen using hydroxypropyl derivatives of beta-cyclodextrin (Hyd-β-CD). The formation of the complex between loxoprofen (LXP) and Hyd-β-CD has been confirmed through changes in absorbance and fluorescence intensity, indicating a 1:1 stoichiometric ratio. The addition of Hyd-β-CD significantly enhanced the solubility of LXP, reflecting successful complexation. Proton NMR and ROESY analyses show that the phenyl group of LXP resides within the cavity of the Hyd-β-CD. Further validation is provided by DFT orientations and their associated energy parameters. Cell viability studies indicate that the increased solubility of the inclusion complex (IC) improves the bioavailability of the drug, while toxicity tests show that our IC material does not induce toxicity in normal cells. Moreover, the anti-inflammatory efficacy of LXP is markedly enhanced in its IC form. These findings underscore the potential of IC in extending the application of LXP, especially in pharmaceuticals, where the enhanced stability and efficacy of natural active ingredients and anti-inflammatory agents are crucial.

Metolachlor Detection in Grain Using N‐Doped Carbon Quantum Dots and the Intramolecular Charge Transfer Effect

Herein, nitrogen is doped into carbon quantum dots (N‐CQDs) using a hydrothermal method for the rapid detection of metolachlor in grain. The morphological features, elemental compositions, and optical properties of the N‐CQDs are then analyzed and investigated using transmission electron microscopy, X‐ray photoelectron spectroscopy, and fluorescence spectroscopy, respectively. Based on the principle of intramolecular charge transfer, a fluorescent probe is constructed for the rapid detection of metolachlor. Under optimal experimental conditions, the fluorescence intensity change values of the N‐CQDs and metolachlor concentration have a good linear relationship when the concentration of metolachlor is in the range of 0.0125−2.5 μg mL−1. An evaluation of the method shows that the method has good selectivity, reproducibility, and stability, with a limit of detection of 1.63 μg kg−1 and a limit of quantification of 3.92 μg kg−1. The spiked recoveries of six real samples are tested using a spiked recovery assay that yielded spiked recoveries in the range of 105.05−87.13%, and their relative standard deviations (n = 3) ranged from 4.62% to 0.61%, indicating that the method can be used for detection in actual samples with good precision and stability.

Lateral flow assay for simultaneous detection of multiple mycotoxins using nanozyme to amplify signals

Co-contamination of multiple mycotoxins produces synergistic toxic effects, leading to more serious hazards. Therefore, the simple, rapid and accurate simultaneous detection of multiple mycotoxins is crucial. Herein, a three-channel aptamer-based lateral flow assay (Apt-LFA) was established for the detection of aflatoxin M1 (AFM1), aflatoxin B1 (AFB1) and ochratoxin A (OTA). The multi-channel Apt-LFA utilized gold‑iridium nanozyme to catalyze the chromogenic substrate, which effectively achieved signal amplification. Moreover, the positions and lengths of the complementary sequences were screened by changes in fluorescence intensity. After grayscale analysis, the semi-quantitative results showed that the detection limits of AFM1, AFB1 and OTA were 0.39 ng/mL, 0.36 ng/mL and 0.82 ng/mL. The recoveries of the multiplexed competitive sensors in complex matrices of real samples were 93.33%–97.01%, 95.72%–102.67%, and 106.88%–109.33%, respectively. In conclusion, the assembly principle of the three-channel Apt-LFA is simple, which can provide a new idea for the simultaneous detection of small molecule targets.

Construction of a monoclonal molecular imprinted sensor with high affinity for specific recognition of influenza a virus subtype

Although molecular imprinting technology has been widely used in the construction of virus sensors, it is still a great challenge to identify subtypes viruses specifically because of their high similarity in morphology, size and structure. Here, a monoclonal molecular imprinted polymers (MIPs) sensor for recognition of H5N1 is constructed to permit the accurate distinguishing of H5N1 from other influenza A virus (IAV) subtypes. Firstly, H5N1 are immobilized on magnetic microspheres to produce H5N1-MagNPs, then the high affinity nanogel H5N1-MIPs is prepared by solid phase imprinting technique. When H5N1-MIPs is combined with MagNP-H5N1, different concentrations of H5N1 are added for competitive substitution. The quantitative detection of H5N1 is realized by the change of fluorescence intensity of supernatant. As expected, the constructed sensor shows satisfactory selectivity, and can identify the target virus from highly similar IAV subtypes, such as H1N1, H7N9 and H9N2. The sensor was highly sensitive, with a detection limit of 0.58 fM, and a selectivity factor that is comparable to that of other small MIPs sensors is achieved. In addition, the proposed sensor is cheap, with a cost of only RMB 0.08 yuan. The proposed monoclonal sensor provides a new method for the specific recognition of designated virus subtype, which is expected to be used for large-scale screening and accurate treatment of infected people.

Facile construction of recyclable heat‐resistant polymers via alkaline‐induced cation‐π cross‐linking

AbstractThe cation‐π cross‐linked heat‐resistant K‐poly(N‐aryleneindole sulphone) PINSU film, which has a high glass transition temperature (Tg = 268°C) and a decomposition temperature (Td = 547°C), was prepared by soaking PINSU film in alkaline aqueous solutions. Compared with the linear polymer PINSU, the Tg and Td of the K‐PINSU were significantly enhanced. The dynamic cation‐π cross‐linking can be reversibly removed and reconstructed driven by external stimulations in the polymer network, which endows the cross‐linked K‐PINSU with cyclable behavior and excellent mechanical stability. In addition, we have successfully demonstrated that the Tg of the K‐PINSU can be nondestructively detected by monitoring the changes in fluorescence intensity.

Exploring the potential of Lactocaseibacillus rhamnosus PMC203 in inducing autophagy to reduce the burden of Mycobacterium tuberculosis

Mycobacterium tuberculosis, a lethal pathogen in human history, causes millions of deaths annually, which demands the development of new concepts of drugs. Considering this fact, earlier research has explored the anti-tuberculosis potential of a probiotic strain, Lactocaseibacillus rhamnosus PMC203, leading to a subsequent focus on the molecular mechanism involved in its effect, particularly on autophagy. In this current study, immunoblotting-based assay exhibited a remarkable expression of autophagy marker LC3-II in the PMC203 treated group compared to an untreated group. A remarkable degradation of p62 was also noticed within treated cells compared to control. Furthermore, the immunofluorescence-based assay showed significant fold change in fluorescence intensity for alexa-647-LC3 and alexa-488-LC3, whereas p62 was degraded noticeably. Moreover, lysosomal biogenesis generation was elevated significantly in terms of LAMP1 and acidic vesicular organelles. As a result, PMC203-induced autophagy played a vital role in reducing M. tuberculosis burden within the macrophages in treated groups compared to untreated group. A colony -forming unit assay also revealed a significant reduction in M. tuberculosis in the treated cells over time. Additionally, the candidate strain significantly upregulated the expression of autophagy induction and lysosomal biogenesis genes. Together, these results could enrich our current knowledge of probiotics-mediated autophagy in tuberculosis and suggest its implications for innovatively managing tuberculosis.

Disinfection-residual bacteria (DRB) after chlorine dioxide treatment: Microbial community structure, regrowth potential, and secretion characteristics

This study investigates the effects of chlorine dioxide (ClO2) disinfection on the community structure, regrowth potential, and metabolic product secretion of disinfection-residual bacteria (DRB) in secondary effluent (SE), denitrification filter effluent (DFE), and ultrafiltration effluent (UE). Results show that ClO2 effectively reduces bacteria in SE and UE, achieving log removal values exceeding 3 at 1 mg/L within 30 min. A salient positive correlation (R2 > 0.95) exists between changes in total fluorescence intensity and disinfection efficacy. Post-treatment, Acinetobacter abundance increased in SE, while Pseudomonas decreased in DFE and UE. At lower ClO2 concentrations, Staphylococcus, Mycobacterium, Aeromonas, and Lactobacillus increased in DFE, but decreased at higher concentrations. After storage, bacterial counts in disinfected samples exceeded those in the control group, surpassing 105 CFU/mL. Despite an initial decline, species richness and evenness partially recovered but remained lower than control levels. Culturing DRB for 72 h showed elevated extracellular polymeric substances (EPS) secretion, quantified as total organic carbon (TOC), ranging from 5 to 27 mg/L, with significantly higher EPS in the disinfection group. Parallel factor analysis with self-organizing maps (PARAFAC-SOM) effectively differentiated water sample types and EPS fluorescent substances, underscoring the potential of three-dimensional fluorescence as an indirect measure of ClO2 disinfection efficacy.

Unravelling structure evolution of dissolved organic matter during oxidation by persulfate: Insights from aromaticity and fluorescence analysis

Persulfate advanced oxidation technology is widely utilized for remediating organic-contaminated groundwater. Post-remediation by persulfate oxidation, the aromaticity of dissolved organic matter (DOM) in groundwater is significantly reduced. Nevertheless, the evolution trends of aromaticity and related structural changes in DOM remained unclear. Here, we selected eight types of DOM to analyze the variation in aromaticity, molecular weight, and fluorescence characteristics during oxidation by persulfate using optical spectroscopy and parallel faction analysis combined with two-dimensional correlation spectroscopy analysis (2D PARAFAC COS). The results showed diverse trends in the changes of aromaticity and maximum fluorescence intensity (Fmax) among different types of DOM as the reaction time increases. Four types of DOM (humic acid 1S104H, fulvic acid, and natural organic matters) exhibited an initially noteworthy increase in aromaticity followed by a decrease, while others demonstrated a continuous decreasing trend (14.3%–69.4%). The overall decreasing magnitude of DOM aromaticity follows the order of natural organic matters ≈ commercial humic acid > fulvic acid > extracted humic acid. The Fmax of humic acid increased, exception of commercial humic acid. The Fmax of fulvic acid initially decreased and then increased, while that of natural organic matters exhibited a decreasing trend (86.4%). The fulvic acid-like substance is the main controlling factor for the aromaticity and molecular weight of DOM during persulfate oxidation process. The oxidation sequence of fluorophores in DOM is as follows: fulvic-like substance, microbial-derived humic-like substance, humic-like substance, and aquatic humic-like substance. The fulvic-like and microbial-derived humic-like substances at longer excitation wavelengths were more sensitive to the response of persulfate oxidation than that of shorter excitation wavelengths. This result reveals the structure evolution of DOM during persulfate oxidation process and provides further support for predicting its environmental behavior.

A rapid method to monitor structural perturbations of high-concentrated therapeutic antibody solutions using Intrinsic Tryptophan Fluorescence Emission spectroscopy

Drug product development of therapeutic antibody formulations is still dictated by the risk of protein particle formation during processing or storage, which can lead to loss of potency and potential immunogenic reactions. Since structural perturbations are the main driver for irreversible protein aggregation, the conformational integrity of antibodies should be closely monitored. The present study evaluated the applicability of a plate reader-based high throughput method for Intrinsic Tryptophan Fluorescence Emission (ITFE) spectroscopy to detect protein aggregation due to protein unfolding in high-concentrated therapeutic antibody samples. The impact of fluorophore concentration on the ITFE signal in microplate readers was investigated by analysis of dilution series of two therapeutic antibodies and pure tryptophan. At low antibody concentrations (< 5 mg/mL, equivalent to 0.8 mM tryptophan), the low inner filter effect suggests a quasi-linear relationship between antibody concentration and ITFE intensity. In contrast, the constant ITFE intensity at high protein concentrations (> 40 mg/mL, equivalent to 6.1 mM tryptophan) indicate that ITFE spectroscopy measurements of IgG1 antibodies are feasible in therapeutically relevant concentrations (up to 223 mg/mL). Furthermore, the capability of the method to detect low levels of unfolding (around 1 %) was confirmed by limit of detection (LOD) determination with temperature-stressed antibody samples as degradation standards. Change of fluorescence intensity at the maximum (ΔIaM) was identified as sensitive descriptor for protein degradation, providing the lowest LOD values. The results demonstrate that ITFE spectroscopy performed in a microplate reader is a valuable tool for high-throughput monitoring of protein degradation in therapeutic antibody formulations.

Substitution Effects on Subcellular Organelle Localization in Live-cell Imaging.

A series of D-p-A indole-containing fluorescent probes were developed followed by an investigation of their photophysical properties and compounds' suitability for subcellular imaging in living cells. We demonstrate that the preference for mitochondrial localization was lost when morpholine was substituted, resulting in the accumulation of the molecule in the lysosomes. However, interestingly, the presence of a nitro group led to their localization within the lipid droplets despite the presence of the morpholine pendant. We also showcase the probes' sensitivity to pH, the influence of added chloroquine, and the temperature response on the changes in fluorescence intensity within lysosomes. The design of the probes with strong intramolecular charge transfer and substantial Stokes shift could facilitate extensive application in various cellular lysosomal models and contribute to a better understanding of the mechanisms involved in stimuli-responsive diseases.

CRASH2p: Closed-loop Two Photon Imaging in Freely Moving Animals.

Direct measurement of neural activity in freely moving animals is essential for understanding how the brain controls and represents behaviors. Genetically encoded calcium indicators report neural activity as changes in fluorescence intensity, but brain motion confounds quantitative measurement of fluorescence. Translation, rotation, and deformation of the brain and the movements of intervening scattering or auto-fluorescent tissue all alter the amount of fluorescent light captured by a microscope. Compared to single-photon approaches, two photon microscopy is less sensitive to scattering and off-target fluorescence, but more sensitive to motion, and two photon imaging has always required anchoring the microscope to the brain. We developed a closed-loop resonant axial-scanning high-speed two photon (CRASH2p) microscope for real-time 3D motion correction in unrestrained animals, without implantation of reference markers. We complemented CRASH2p with a novel scanning strategy and a multi-stage registration pipeline. We performed volumetric ratiometrically corrected functional imaging in the CNS of freely moving Drosophila larvae and discovered previously unknown neural correlates of behavior.

CD52/FLAG and CD52/HA Fusion Proteins as Novel Magnetic Cell Selection Markers.

At present, the magnetic selection of genetically modified cells is mainly performed with surface markers naturally expressed by cells such as CD4, LNGFR (low affinity nerve growth factor receptor), and MHC class I molecule H-2Kk. The disadvantage of such markers is the possibility of their undesired and poorly predictable expression by unmodified cells before or after cell manipulation, which makes it essential to develop new surface markers that would not have such a drawback. Earlier, modified CD52 surface protein variants with embedded HA and FLAG epitope tags (CD52/FLAG and CD52/HA) were developed by the group of Dr. Mazurov for the fluorescent cell sorting of CRISPR-modified cells. In the current study, we tested whether these markers can be used for the magnetic selection of transduced cells. For this purpose, appropriate constructs were created in MigR1-based bicistronic retroviral vectors containing EGFP and DsRedExpress2 as fluorescent reporters. Cytometric analysis of the transduced NIH 3T3 cell populations after magnetic selection evaluated the efficiency of isolation and purity of the obtained populations, as well as the change in the median fluorescence intensity (MFI). The results of this study demonstrate that the surface markers CD52/FLAG and CD52/HA can be effectively used for magnetic cell selection, and their efficiencies are comparable to that of the commonly used LNGFR marker. At the same time, the significant advantage of these markers is the absence of HA and FLAG epitope sequences in cellular proteins, which rules out the spurious co-isolation of negative cells.

A fluorescent probe for specific dual recognition of Ni2+ and pH

Nickel ion (Ni2+) and pH play an important role in environment and living organisms. A fluorescent probe “naphthalimide- s-triazine” (NCNS) for targeted dual detection of Ni2+ and pH was synthesized. As a result, NCNS exhibits excellent optical properties: a much larger Stokes shift (140 nm), eminent changes of fluorescence intensity and significant red-shift both for Ni2+ and pH. As for the detection of Ni2+, the selectivity is high and the anti-interference is strong. NCNS can fluorescently detect Ni2+ in a wider pH range from 4.0 to 10.5. It provides a much lower limit of detection (LOD, 20.03 nM), a rapid response time (150 s) and six times reversibility, showing the high sensitivity. Particularly, NCNS can be applied to fluorescently detect Ni2+ in actual water samples and HA-VSMC imaging. In the detection of pH, the probe generates a ratiometric fluorescence in a wide pH range (3.0 ∼ 12.3). NCNS has been successfully made test paper both for Ni2+ and pH. The mechanisms of the double recognition are verified by the density functional theory (DFT) calculations and the nuclear magnetic resonance (NMR) titration experiments.

Rapid detection and quantification of milk adulterants using a nanoclusters-based fluorescent optical tongue

A paper-based sensor array consisting of eight nanoclusters (NCs) combined with multivariate analysis was used as a rapid method for the determination of animal sources of milk; goat, camel, sheep and cow. It was also used to detect and quantify three adulterants including sodium hypochlorite, hydrogen peroxide and formaldehyde in milk. The changes in fluorescence intensity of the NCs were quantified using a smartphone when the sensor array was immersed in the milk samples. The device generated a specific colorimetric signature for milk samples from different animals and for different adulterants. This allowed simultaneous identification of animal and adulterant sources with 100% accuracy. The device was found to be capable of accurately measuring the level of contaminants with a detection limit as low as 0.01% using partial least squares regression. In conclusion, a paper-based optical tongue device has been developed for the detection of adulterants in milk with point-of-need capability.

Aptamer-carbon quantum dots and silver nanoparticles construct a FRET sensor for sensitive detection of E. coli

E. coli is considered to be one of the most dangerous foodborne pathogens and precise detection of E. coli in food is required to ensure the quality and safety of food. In this work, carbon quantum dots (CQDs) were prepared from grapefruit peel as a carbon source by microwave heating method. A fluorescent resonance energy transfer (FRET)-based fluorescent biosensor was constructed by combining CQDs with silver nanoparticles (Ag NPs) for rapid detection of E. coli (BL21). The prepared CQDs showed good dispersion under transmission electron microscopy (TEM), with a spherical shape and an average particle size of 7.4 nm. The CQDs showed bright green fluorescence under UV light irradiation, which was strongly fluorescent (> 1.0 × 103 a.u.) and there was no change in fluorescence intensity over 150 days. Its fluorescence transduction is based on the spectral overlap between the donor (CQDs) emission and the acceptor (nanoparticles) absorbance. The fluorescence of CQDs attached to the aptamer is burst in the presence of Ag NPs. Upon addition of a specific E. coli (BL21) solution, an aptamer-target complex is formed and the preferential interaction of the aptamer with the specific bacteria leads to the release of CQDs and Ag NPs. After a period of incubation, the bacteria are centrifuged, resulting in the precipitation of E. coli aptamer couples and Ag NPs, and therefore the recovery of CQDs fluorescence. This method allows specific detection of E. coli (BL21) in a wide range of pathogenic bacteria. The final results show that the sensor has a linear range of 2×103 ∼ 2×108 CFU·mL−1 and a low detection limit of 77 CFU·mL−1 for E. coli.

Synthesis and Characterization of Multiple Stimuli-Responsive Fluorescent Polymer Hydrogels Based on Terpyridine and N-Isopropylacrylamide.

A series of stimuli-responsive fluorescent hydrogels were successfully synthesized via micelle radical copolymerization of hydrophilic acrylamide (AM), hydrophobic chromophore terpyridine-based monomer (TPY), and N-isopropylacrylamide (NIPAM). These hydrogels presented blue emissions (423-440 nm) under room temperature, which is caused by the π-π* transition of the conjugated structures. Once the ambient temperature was increased to 55 °C, the fluorescence color changed from blue (430 nm) to pink (575 nm) within 10 min, subsequently to yellow (535 nm), and eventually back to pink. The thermal-responsive properties are attributed to the transition of the TPY units from unimer to dimer aggregation via the intermolecular charge transfer complex at high temperatures. The hydrogels showed pH-responsive properties. The emission peak of the hydrogel exhibited a blue shift of ~54 nm from neuter conditions to acidic conditions, while a 6 nm red shift to an alkaline environment was observed. The hydrogels demonstrated an obvious change in fluorescence intensity and wavelength upon adding different metal ions, which is caused by the coordination between the terpyridine units incorporated on the backbones and the metal ions. As a consequence, the hydrogels presented a sharp quenching fluorescence interaction with Fe2+, Fe3+, Cu2+, Hg2+, Ni2+, and Co2+, while it exhibited an enhanced fluorescence intensity interaction with Sn2+, Cd2+, and Zn2+. The microstructural, mechanical, and rheological properties of these luminescent hydrogels have been systematically investigated.