Photosystem II monomeric antenna CP26 plays a key role in nonphotochemical quenching in Chlamydomonas.

Thermal dissipation of excess excitation energy, called nonphotochemical quenching (NPQ), is 1 of the main photoprotective mechanisms in oxygenic photosynthetic organisms. Here, we investigated the function of the monomeric photosystem II (PSII) antenna protein CP26 in photoprotection and light harvesting in Chlamydomonas reinhardtii, a model organism for green algae. We used CRISPR/Cas9 genome editing and complementation to generate cp26 knockout mutants (named k6#) that did not negatively affect CP29 accumulation, which differed from previous cp26 mutants, allowing us to compare mutants specifically deprived of CP26, CP29, or both. The absence of CP26 partially affected PSII activity, causing reduced growth at low or medium light but not at high irradiances. However, the main phenotype observed in k6# mutants was a more than 70% reduction of NPQ compared to the wild type (Wt). This phenotype was fully rescued by genetic complementation and complemented strains accumulating different levels of CP26, demonstrating that ∼50% of CP26 content, compared to the Wt, was sufficient to restore the NPQ capacity. Our findings demonstrate a pivotal role for CP26 in NPQ induction, while CP29 is crucial for PSII activity. The genetic engineering of these 2 proteins could be a promising strategy to regulate the photosynthetic efficiency of microalgae under different light regimes.

Enhancement of Photoluminescence Properties via Polymer Infiltration in a Colloidal Photonic Glass.

Photonic glasses (PGs) based on the self-assembly of monosized nanoparticles can be an effective tool for realizing disordered structures capable of tailoring light diffusion due to the establishment of Mie resonances. In particular, the wavelength position of these resonances depends mainly on the morphology (dimension) and optical properties (refractive index) of the building blocks. In this study, we report the fabrication and optical characterization of photonic glasses obtained via a self-assembling technique. Furthermore, we have demonstrated that the infiltration of these systems with a green-emitting polymer enhances the properties of the polymer, resulting in a large increase in its photoluminescence quantum yield and a 3 ps growing time of the photoluminescence time decay Finally, the development of the aforementioned system can serve as a suitable low-cost platform for the realization of lasers and fluorescence-based bio-sensors.

Dynamics of Two Distinct Exciton Populations in Methyl-Functionalized Germanane.

Methyl-substituted germanane is an emerging material that has been proposed for novel applications in optoelectronics, photoelectrocatalysis, and biosensors. It is a two-dimensional semiconductor with a strong above-gap fluorescence associated with water intercalation. Here, we use time-resolved photoluminescence spectroscopy to understand the mechanism causing this fluorescence. We show that it originates from two distinct exciton populations. Both populations recombine exponentially, accompanied by the thermally activated transfer of exciton population from the shorter- to the longer-lived type. The two exciton populations involve different electronic levels and couple to different phonons. The longer-lived type of exciton migrates within the disordered energy landscape of localized recombination centers. These outcomes shed light on the fundamental optical and electronic properties of functionalized germanane, enabling the groundwork for future applications in optoelectronics, light harvesting, and sensing.

Azobenzene photoisomerization probes cell membrane viscosity.

The viscosity of cell membranes is a crucial parameter that affects the diffusion of small molecules both across and within the lipid membrane and that is related to several diseases. Therefore, the possibility to measure quantitatively membrane viscosity on the nanoscale is of great interest. Here, we report a complete investigation of the photophysics of an amphiphilic membrane-targeted azobenzene (ZIAPIN2) and we propose its use as a viscosity probe for cell membranes. We exploit ZIAPIN2 trans-cis photoisomerization to develop a molecular viscometer and to assess the viscosity of Escherichia coli bacteria membranes employing time-resolved fluorescence spectroscopy. Fluorescence lifetime measurements of ZIAPIN2 in E. coli bacteria suspensions correctly indicate that the membrane viscosity decreases as the temperature of the sample increases. Given the non-homogeneity and the anisotropy of cell membranes, as supported by the photophysical characterization of the probe within the lipid bilayer, we shed new light on the intricate membrane rheology.

Molecular mechanisms of light harvesting in the minor antenna CP29 in near-native membrane lipidic environment.

CP29, a chlorophyll a/b-xanthophyll binding protein, bridges energy transfer between the major LHCII antenna complexes and photosystem II reaction centers. It hosts one of the two identified quenching sites, making it crucial for regulated photoprotection mechanisms. Until now, the photophysics of CP29 has been studied on the purified protein in detergent solutions since spectrally overlapping signals affect in vivo measurements. However, the protein in detergent assumes non-native conformations compared to its physiological state in the thylakoid membrane. Here, we report a detailed photophysical study on CP29 inserted in discoidal lipid bilayers, known as nanodiscs, which mimic the native membrane environment. Using picosecond time-resolved fluorescence and femtosecond transient absorption (TA), we observed shortening of the Chl fluorescence lifetime with a decrease of the carotenoid triplet formation yield for CP29 in nanodiscs as compared to the protein in detergent. Global analysis of TA data suggests a 1Chl* quenching mechanism dependent on excitation energy transfer to a carotenoid dark state, likely the proposed S*, which is believed to be formed due to a carotenoid conformational change affecting the S1 state. We suggest that the accessibility of the S* state in different local environments plays a key role in determining the quenching of Chl excited states. In vivo, non-photochemical quenching is activated by de-epoxidation of violaxanthin into zeaxanthin. CP29-zeaxanthin in nanodiscs further shortens the Chl lifetime, which underlines the critical role of zeaxanthin in modulating photoprotection activity.

Noniridescent Biomimetic Photonic Microdomes by Inkjet Printing.

Certain bird species have evolved spectacular colors that arise from organized nanostructures of melanin. Its high refractive index (∼1.8) and broadband absorptive properties enable vivid structural colors that are nonsusceptible to photobleaching. Mimicking natural melanin structural coloration could enable several important applications, in particular, for noniridescent systems with colors that are independent of incidence angle. Here, we address this by forming melanin photonic crystal microdomes by inkjet printing. Owing to their curved nature, the microdomes exhibit noniridescent vivid structural coloration, tunable throughout the visible range via the size of the nanoparticles. Large-area arrays (>1 cm2) of high-quality photonic microdomes could be printed on both rigid and flexible substrates. Combined with scalable fabrication and the nontoxicity of melanin, the presented photonic microdomes with noniridescent structural coloration may find use in a variety of applications, including sensing, displays, and anticounterfeit holograms.

Ultrafast dynamics in co-sensitized photocatalysts under visible and NIR light irradiation.

Co-sensitization to achieve a broad absorption window is a widely accepted technique in light harvesting nanohybrid synthesis. Protoporphyrin (PPIX) and squaraine (SQ2) are two organic sensitizers absorbing in the visible and NIR wavelength regions of the solar spectrum, respectively. In the present study, we have sensitized zinc oxide (ZnO) nanoparticles using PPIX and SQ2 simultaneously for their potential use in broad-band solar light harvesting in photocatalysis. Förster resonance energy transfer (FRET) from PPIX to SQ2 in close proximity to the ZnO surface has been found to enhance visible light photocatalysis. In order to confirm the effect of intermolecular FRET in photocatalysis, the excited state lifetime of the energy donor dye PPIX has been modulated by inserting d10 (ZnII) and d7 (CoII) metal ions in the central position of the dye (PP(Zn) and PP(Co)). In the case of PP(Co)-SQ2, extensive photo-induced ligand to metal charge transfer counteracts the FRET efficiency while efficient FRET has been observed for the PP(Zn)-SQ2 pair. This observation has been justified by the comparison of the visible light photocatalysis of the respective nanohybrids with several control studies. We have also investigated the NIR photocatalysis of the co-sensitized nanohybrids which reveals that reduced aggregation of SQ2 due to co-sensitization of PPIX increases the NIR photocatalysis. However, core-metalation of PPIX reduces the NIR photocatalytic efficacy, most probably due to excited state charge transfer from SQ2 to the metal centre of PP(Co)/PP(Zn) through the conduction band of the host ZnO nanoparticles.

Sensitized ZnO nanorod assemblies to detect heavy metal contaminated phytomedicines: spectroscopic and simulation studies.

The immense pharmacological relevance of the herbal medicine curcumin including anti-cancer and anti-Alzheimer effects, suggests it to be a superior alternative to synthesised drugs. The diverse functionalities with minimal side effects intensify the use of curcumin not only as a dietary supplement but also as a therapeutic agent. Besides all this effectiveness, some recent literature reported the presence of deleterious heavy metal contaminants from various sources in curcumin leading to potential health hazards. In this regard, we attempt to fabricate ZnO based nanoprobes to detect metal conjugated curcumin. We have synthesized and structurally characterized the ZnO nanorods (NR). Three samples namely curcumin (pure), Zn-curcumin (non-toxic metal attached to curcumin) and Hg-curcumin (toxic heavy metal attached to curcumin) were prepared for consideration. The samples were electrochemically deposited onto ZnO surfaces and the attachment was confirmed by cyclic voltammetry experiments. Moreover, to confirm a molecular level interaction picosecond-resolved PL-quenching of ZnO NR due to Förster Resonance Energy Transfer (FRET) from donor ZnO NR to the acceptor curcumin moieties was employed. The attachment proximity of ZnO NR and curcumin moieties depends on the size of metals. First principles analysis suggests a variance of attachment sites and heavy metal Hg conjugated curcumin binds through a peripheral hydroxy group to NR. We fabricated a facile photovoltaic device consisting of ZnO NR as the working electrode with Pt counter electrode and iodide-triiodide as the electrolyte. The trend in photocurrent under visible light illumination suggests an enhancement in the case of heavy metal ions due to long range interaction and greater accumulation of charge at the active electrode. Our results provide a detailed physical insight into interfacial processes that are crucial for detecting heavy-metal attached phytomedicines and are thus expected to find vast application as sensors for the detection of selective metal contaminants.

Photoinduced Dynamics and Toxicity of a Cancer Drug in Proximity of Inorganic Nanoparticles under Visible Light.

Drug sensitization with various inorganic nanoparticles (NPs) has proved to be a promising and an emergent concept in the field of nanomedicine. Rose bengal (RB), a notable photosensitizer, triggers the formation of reactive oxygen species under green-light irradiation, and consequently, it induces cytotoxicity and cell death. In the present study, the effect of photoinduced dynamics of RB upon complexation with semiconductor zinc oxide NPs is explored. To accomplish this, we successfully synthesized nanohybrids of RB with ZnO NPs with a particle size of 24 nm and optically characterized them. The uniform size and integrity of the particles were confirmed by high-resolution transmission electron microscopy. UV/Vis absorption and steady-state fluorescence studies reveal the formation of the nanohybrids. ultrafast picosecond-resolved fluorescence studies of RB-ZnO nanohybrids demonstrate an efficient electron transfer from the photoexcited drug to the semiconductor NPs. Picosecond-resolved Förster resonance energy transfer from ZnO NPs to RB unravel the proximity of the drug to the semiconductor at the molecular level. The photoinduced ROS formation was monitored using a dichlorofluorescin oxidation assay, which is a conventional oxidative stress indicator. It is observed that the ROS generation under green light illumination is greater at low concentrations of RB-ZnO nanohybrids compared with free RB. Substantial photodynamic activity of the nanohybrids in bacterial and fungal cell lines validated the in vitro toxicity results. Furthermore, the cytotoxic effect of the nanohybrids in HeLa cells, which was monitored by MTT assay, is also noteworthy.

Heterodimerization at the dye sensitized TiO2 surface: an efficient strategy toward quick removal of water contaminants.

Sensitization of wide bandgap semiconductors with heterodimers for better solar light sensitivity has attracted widespread attention in the recent times. However, application of heterodimerization for removing soluble water pollutants from waste water is sparse in the literature. In the present study, we have utilized heterodimerization of a model pollutant methylene blue (MB) with a ruthenium based dye N719 for the removal of the pollutant. We have synthesized N719 functionalized carbonate doped TiO2 microspheres (doped MS) which act as a novel material for the detoxification of MB containing water by adsorbing at the surface and eventually killing by photoinduced reduction under visible light irradiation. The mechanism of surface adsorption and photoreduction of MB are explored using steady state and time resolved spectroscopy studies. We have fabricated two types of prototype devices (flow device and active filter) using the functionalized doped MS. Both the devices show excellent dye removal activity and recyclability. The present study would find relevance in the removal of soluble pollutants from waste water.

Facile synthesis of reduced graphene oxide-gold nanohybrid for potential use in industrial waste-water treatment.

Here, we report a facile approach, by the photochemical reduction technique, for in situ synthesis of Au-reduced graphene oxide (Au-RGO) nanohybrids, which demonstrate excellent adsorption capacities and recyclability for a broad range of dyes. High-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) data confirm the successful synthesis of Au-RGO nanohybrids. The effect of several experimental parameters (temperature and pH) variation can effectively control the dye adsorption capability. Furthermore, kinetic adsorption data reveal that the adsorption process follows a pseudo second-order model. The negative value of Gibbs free energy (ΔG0) confirms spontaneity while the positive enthalpy (ΔH0) indicates the endothermic nature of the adsorption process. Picosecond resolved fluorescence technique unravels the excited state dynamical processes of dye molecules adsorbed on the Au-RGO surface. Time resolved fluorescence quenching of Rh123 after adsorption on Au-RGO nanohybrids indicates efficient energy transfer from Rh123 to Au nanoparticles. A prototype device has been fabricated using Au-RGO nanohybrids on a syringe filter (pore size: 0.220 µm) and the experimental data indicate efficient removal of dyes from waste water with high recyclability. The application of this nanohybrid may lead to the development of an efficient reusable adsorbent in portable water purification.

Direct observation of key photoinduced dynamics in a potential nano-delivery vehicle of cancer drugs.

In recent times, significant achievements in the use of zinc oxide (ZnO) nanoparticles (NPs) as delivery vehicles of cancer drugs have been made. The present study is an attempt to explore the key photoinduced dynamics in ZnO NPs upon complexation with a model cancer drug protoporphyrin IX (PP). The nanohybrid has been characterized by FTIR, Raman scattering and UV-Vis absorption spectroscopy. Picosecond-resolved Förster resonance energy transfer (FRET) from the defect mediated emission of ZnO NPs to PP has been used to study the formation of the nanohybrid at the molecular level. Picosecond-resolved fluorescence studies of PP-ZnO nanohybrids reveal efficient electron migration from photoexcited PP to ZnO, eventually enhancing the ROS activity. The dichlorofluorescin (DCFH) oxidation and no oxidation of luminol in PP/PP-ZnO nanohybrids upon green light illumination unravel that the nature of ROS is essentially singlet oxygen rather than superoxide anions. Surface mediated photocatalysis of methylene blue (MB) in an aqueous solution of the nanohybrid has also been investigated. Direct evidence of the role of electron transfer as a key player in enhanced ROS generation from the nanohybrid is also clear from the photocurrent measurement studies. We have also used the nanohybrid in a model photodynamic therapy application in a light sensitized bacteriological culture experiment.

Sensitization of an endogenous photosensitizer: electronic spectroscopy of riboflavin in the proximity of semiconductor, insulator, and metal nanoparticles.

Riboflavin (Rf) is a class of important vitamins (Vitamin B2) and a well-known antioxidant. Here we have synthesized nanohybrids of Rf with a number of inorganic nanoparticles (NPs); namely zinc oxide (ZnO), titanium oxide (TiO2), aluminum oxide (Al2O3) and gold NPs of similar sizes. While high resolution transmission electron microscopy (HRTEM) confirms integrity and sizes of the NPs, intactness of the molecular structure of the drug Rf is revealed from absorption and steady-state emission spectra of the drug in the nanohybrid. Raman spectroscopy on the nanohybrids shows the nature of molecular complexation of the drug with the inorganic NPs. For the semiconductor and insulator NPs, the complexation is found to be noncovalent, however, a covalent attachment of the drug with the dangling bonds of metal atoms at the surface is observed. In order to investigate antioxidant activity of the nanohybrids, we have performed 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay of the nanohybrids in dark as well as under blue light irradiation. Whereas change of the antioxidant activity of the nanohybrids with respect to free riboflavin in the absence of light is observed to be insignificant, a drastic change in the activity in the case of TiO2 and ZnO in the presence of light is evident. No change in the case of Al2O3 and a significant decrease in the antioxidant activity for gold nanohybrids are also remarkable. Picosecond-resolved fluorescence studies on the nanohybrids reveal a molecular picture of the differential antioxidant activities. An ultrafast photoinduced electron transfer from Rf to ZnO and TiO2 are clearly evident from the corresponding fluorescence transients. We have compared the picosecond-resolved transients with that of Rf in the presence of a well-known electron acceptor benzoquinone (BQ) and found similar time scales. No temporal change in the fluorescence transient of riboflavin in Al2O3 nanohybrids compared to that of free Rf is observed indicating uneventful excited state relaxation of the nanohybrids. Nanosurface energy transfer (NSET) over Förster resonance energy transfer (FRET) is found to be the prevailing de-excitation mechanism in the case of gold nanohybrids, because of the strong spectral overlap between Rf emission and surface plasmon absorption of the gold NPs. Different excited state mechanisms as revealed from our studies are expected to be useful for the design of NP-sensitized drugs, which are reported sparsely in the literature.

Impact of metal ions in porphyrin-based applied materials for visible-light photocatalysis: key information from ultrafast electronic spectroscopy.

Protoporphyrin IX-zinc oxide (PP-ZnO) nanohybrids have been synthesized for applications in photocatalytic devices. High-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), and steady-state infrared, absorption, and emission spectroscopies have been used to analyze the structural details and optical properties of these nanohybrids. Time-resolved fluorescence and transient absorption techniques have been applied to study the ultrafast dynamic events that are key to photocatalytic activities. The photocatalytic efficiency under visible-light irradiation in the presence of naturally abundant iron(III) and copper(II) ions has been found to be significantly retarded in the former case, but enhanced in the latter case. More importantly, femtosecond (fs) transient absorption data have clearly demonstrated that the residence of photoexcited electrons from the sensitizer PP in the centrally located iron moiety hinders ground-state bleach recovery of the sensitizer, affecting the overall photocatalytic rate of the nanohybrid. The presence of copper(II) ions, on the other hand, offers additional stability against photobleaching and eventually enhances the efficiency of photocatalysis. In addition, we have also explored the role of UV light in the efficiency of photocatalysis and have rationalized our observations from femtosecond- to picosecond-resolved studies.

Probing the Effect of Mutations on Light Harvesting in CP29 by Transient Absorption and First-Principles Simulations.

Natural light harvesting is exceptionally efficient thanks to the local energy funnel created within light-harvesting complexes (LHCs). To understand the design principles underlying energy transport in LHCs, ultrafast spectroscopy is often complemented by mutational studies that introduce perturbations into the excitonic structure of the natural complexes. However, such studies may fall short of identifying all excitation energy transfer (EET) pathways and their changes upon mutation. Here, we show that a synergistic combination of first-principles calculations and ultrafast spectroscopy can give unprecedented insight into the EET pathways occurring within LHCs. We measured the transient absorption spectra of the minor CP29 complex of plants and of two mutants, systematically mapping the kinetic components seen in experiments to the simulated exciton dynamics. With our combined strategy, we show that EET in CP29 is surprisingly robust to the changes in the exciton states induced by mutations, explaining the versatility of plant LHCs.

Influence of structural colour on the photoprotective mechanism in the gametophyte phase of the red alga Chondrus crispus.

Marine life is populated by a huge diversity of organisms with an incredible range of colour. While structural colour mechanisms and functions are usually well studied in marine animal species, there is a huge knowledge gap regarding the marine macroalgae (red, green and brown seaweeds) that have structural coloration and the biological significance of this phenomenon in these photosynthetic organisms. Here we show that structural colour in the gametophyte life history phase of the red alga Chondrus crispus plays an important role as a photoprotective mechanism in synergy with the other pigments present. In particular, we have demonstrated that blue structural coloration attenuates the more energetic light while simultaneously favouring green and red light harvesting through the external antennae (phycobilisomes) which possess an intensity-dependent photoprotection mechanism. These insights into the relationship between structural colour and photosynthetic light management further our understanding of the mechanisms involved.

Site-Directed Mutagenesis of the Chlorophyll-Binding Sites Modulates Excited-State Lifetime and Chlorophyll-Xanthophyll Energy Transfer in the Monomeric Light-Harvesting Complex CP29.

We combine site-directed mutagenesis with picosecond time-resolved fluorescence and femtosecond transient absorption (TA) spectroscopies to identify excitation energy transfer (EET) processes between chlorophylls (Chls) and xanthophylls (Xant) in the minor antenna complex CP29 assembled inside nanodiscs, which result in quenching. When compared to WT CP29, a longer lifetime was observed in the A2 mutant, missing Chl a612, which closely interacts with Xant Lutein in site L1. Conversely, a shorter lifetime was obtained in the A5 mutant, in which the interaction between Chl a603 and Chl a609 is strengthened, shifting absorption to lower energy and enhancing Chl-Xant EET. Global analysis of TA data indicated that EET from Chl a Qy to a Car dark state S* is active in both the A2 and A5 mutants and that their rate constants are modulated by mutations. Our study provides experimental evidence that multiple Chl-Xant interactions are involved in the quenching activity of CP29.

Role of central metal ions in hematoporphyrin-functionalized titania in solar energy conversion dynamics.

In this study, we have investigated the efficacy of electron transfer processes in hematoporphyrin (HP) and iron hematoporphyrin ((Fe)HP) sensitized titania as potential materials for capturing and storing solar energy. Steady-state and picosecond-resolved fluorescence studies show the efficient photoinduced electron transfer processes in hematoporphyrin-TiO2 (HP-TiO2) and Fe(III)-hematoporphyrin-TiO2 (Fe(III)HP-TiO2) nanohybrids, which reveal the role of central metal ions in electron transfer processes. The bidentate covalent attachment of HP onto TiO2 particulates is confirmed by FTIR, Raman scattering and X-ray photoelectron spectroscopy (XPS) studies. The iron oxidation states and the attachment of iron to porphyrin through pyrrole nitrogen atoms were investigated by cyclic voltammetry and FTIR studies, respectively. We also investigated the potential application of HP-TiO2 and Fe(III)HP-TiO2 nanohybrids for the photodegradation of a model organic pollutant methylene blue (MB) in aqueous solution under wavelength dependent light irradiation. To further investigate the role of iron oxidation states in electron transfer processes, photocurrent measurements were done by using Fe(III) and Fe(II) ions in porphyrin. This work demonstrates the role of central metal ions in fundamental electron transfer processes in porphyrin sensitized titania and their implications for dye-sensitized device performance.

Toward Photoactive Wallpapers Based on ZnO-Cellulose Nanocomposites.

The quest for eco-friendly materials with anticipated positive impact for sustainability is crucial to achieve the UN sustainable development goals. Classical strategies of composite materials can be applied on novel nanomaterials and green materials. Besides the actual technology and applications also processing and manufacturing methods should be further advanced to make entire technology concepts sustainable. Here, they show an efficient way to combine two low-cost materials, cellulose and zinc oxide (ZnO), to achieve novel functional and "green" materials via paper-making processes. While cellulose is the most abundant and cost-effective organic material extractable from nature. ZnO is cheap and known of its photocatalytic, antibacterial, and UV absorption properties. ZnO nanowires are grown directly onto cellulose fibers in water solutions and then dewatered in a process mimicking existing steps of large-scale papermaking technology. The ZnO NW paper exhibits excellent photo-conducting properties under simulated sunlight with good ON/OFF switching and long-term stability (90 minutes). It also acts as an efficient photocatalyst for hydrogen peroxide (H2O2) generation (5.7 × 10-9 m s-1) with an envision the possibility of using it in buildings to enable large surfaces to spontaneously produce H2O2 at its outer surface. Such technology promise for fast degradation of microorganisms to suppress the spreading of diseases.

Membrane Order Effect on the Photoresponse of an Organic Transducer.

Non-genetic photostimulation, which allows for control over cellular activity via the use of cell-targeting phototransducers, is widely used nowadays to study and modulate/restore biological functions. This approach relies on non-covalent interactions between the phototransducer and the cell membrane, thus implying that cell conditions and membrane status can dictate the effectiveness of the method. For instance, although immortalized cell lines are traditionally used in photostimulation experiments, it has been demonstrated that the number of passages they undergo is correlated to the worsening of cell conditions. In principle, this could impact cell responsivity against exogenous stressors, including photostimulation. However, these aspects have usually been neglected in previous experiments. In this work, we investigated whether cell passages could affect membrane properties (such as polarity and fluidity). We applied optical spectroscopy and electrophysiological measurements in two different biological models: (i) an epithelial immortalized cell line (HEK-293T cells) and (ii) liposomes. Different numbers of cell passages were compared to a different morphology in the liposome membrane. We demonstrated that cell membranes show a significant decrease in ordered domains upon increasing the passage number. Furthermore, we observed that cell responsivity against external stressors is markedly different between aged and non-aged cells. Firstly, we noted that the thermal-disordering effect that is usually observed in membranes is more evident in aged cells than in non-aged ones. We then set up a photostimulation experiment by using a membrane-targeted azobenzene as a phototransducer (Ziapin2). As an example of a functional consequence of such a condition, we showed that the rate of isomerization of an intramembrane molecular transducer is significantly impaired in aged cells. The reduction in the photoisomerization rate translates in cells with a sustained reduction of the Ziapin2-related hyperpolarization of the membrane potential and an overall increase in the molecule fluorescence. Overall, our results suggest that membrane stimulation strongly depends on membrane order, highlighting the importance of cell passage during the characterization of the stimulation tools. This study can shine light on the correlation between aging and the development of diseases driven by membrane degradation as well as on the different cell responsivities against external stressors, such as temperature and photostimulation.