Resonance Energy Transfer Pair Research Articles

Extremely Sensitive Genetically Encoded Temperature Indicator Enabling Measurement at the Organelle Level.

Intracellular temperature is a fundamental parameter in biochemical reactions. Genetically encoded fluorescent temperature indicators (GETIs) have been developed to visualize intracellular thermogenesis; however, the temperature sensitivity or localization capability in specific organelles should have been further improved to clearly capture when and where intracellular temperature changes at the subcellular level occur. Here, we developed a new GETI, gMELT, composed of donor and acceptor subunits, in which cyan and yellow fluorescent proteins, respectively, as a Förster resonance energy transfer (FRET) pair were fused with temperature-sensitive domains. The donor and acceptor subunits associated and dissociated in response to temperature changes, altering the FRET efficiency. Consequently, gMELT functioned as a fluorescence ratiometric indicator. Untagged gMELT was expressed in the cytoplasm, whereas versions fused with specific localization signals were targeted to the endoplasmic reticulum (ER) or mitochondria. All gMELT variations enabled more sensitive temperature measurements in cellular compartments than those in previous GETIs. The gMELTs, tagged with ER or mitochondrial targeting sequences, were used to detect thermogenesis in organelles stimulated chemically, a method previously known to induce thermogenesis. The observed temperature changes were comparable to previous reports, assuming that the fluorescence readout changes were exclusively due to temperature variations. Furthermore, we demonstrated how macromolecular crowding influences gMELT fluorescence given that this factor can subtly affect the fluorescence readout. Investigating thermogenesis with gMELT, accounting for factors such as macromolecular crowding, will enhance our understanding of intracellular thermogenesis phenomena.

A novel “on-off-on” electrochemiluminescence aptasensor based on resonance energy transfer as signal amplification strategy for ultrasensitive detection of sulfadiazine in different samples

The misuse of antibiotics may contaminate the environment and cause harm to human health. Therefore, rapid and accurate detection of antibiotics is essential. In this study, a novel electrochemiluminescence resonance energy transfer (ECL-RET) pair was designed using a new ECL emitter (CPM, Ce-TBAPy) as the donor and Co-MOF@AuPt as the acceptor. Moreover, a highly sensitive and specific “on-off-on” ECL aptasensor was constructed for detecting sulfadiazine (SDZ). The aptasensor exhibited a broad linear range (from 10.0 fg mL−1 to 100 ng mL−1) for the SDZ concentration, with limit of detection and limit of quantification values of 1.14 fg mL−1and 3.75 fg mL−1, respectively. The aptasensor achieved good results in spiking experiments with milk and egg samples, and successfully quantified SDZ in fish meal quality control sample. The prepared aptasensor presents great potential for food and environmental safety by detecting antibiotics.

Single- and two-photon-induced Förster resonance energy transfer in InP-mCherry bioconjugates.

Indium phosphide (InP) quantum dots (QDs) have recently garnered considerable interest in the design of bioprobes due to their non-toxic nature and excellent optical properties. Several attempts for the conjunction of InP QDs with various entities such as organic dyes and dye-labeled proteins have been reported, while that with fluorescent proteins remains largely uncharted. This study reports the development of a Förster resonance energy transfer pair comprising glutathione-capped InP/GaP/ZnS QDs [InP(G)] and the fluorescent protein mCherry. Glutathione on InP(G) undergoes effective bioconjugation with mCherry consisting of a hexahistidine tag, and the nonradiative energy transfer is investigated using steady-state and time-resolved measurements. Selective one-photon excitation of InP(G) in the presence of mCherry shows a decay of the emission of the QDs and a concomitant growth of acceptor emission. Time-resolved investigations prove the nonradiative transfer of energy between InP(G) and mCherry. Furthermore, the scope of two-photon-induced energy transfer between InP(G) and mCherry is investigated by exciting the donor in the optical transparency range. The two-photon absorption is confirmed by the quadratic relationship between the emission intensity and the excitation power. In general, near-infrared excitation provides a path for effective light penetration into the tissues and reduces the photodamage of the sample. The two-photon-induced energy transfer in such assemblies could set the stage for a wide range of biological and optoelectronic applications in the foreseeable future.

Amphiphilic Poly(2-oxazoline)s with Glycine-Containing Hydrophobic Blocks Tailored for Panobinostat- and Imatinib-Loaded Micelles.

Aiming toward the development of tailored carrier materials for the cytostatics panobinostat and imatinib, an amphiphilic block copolymer composed of poly(2-ethyl-2-oxazoline) and a degradable poly(2-(3-phenylpropyl)-2-oxazoline) analogue (dPPhPrOx-b-PEtOx) was synthesized via a postpolymerization synthesis route based on reacylation of oxidized linear poly(ethylene imine). The obtained dPPhPrOx-b-PEtOx was found to readily self-assemble into well-defined micelles with a critical micelle concentration of 1μgmL-1. The incubation of HUVEC cells with the blank micelles revealed their excellent cytocompatibility (up to 2 mg mL-1), thus confirming the polymers' suitability for potential drug delivery application. Subsequently, the encapsulation of the two cytostatics, panobinostat and imatinib, into the dPPhPrOx-b-PEtOx micelles was successfully demonstrated (Dh ≈ 80 nm, PDI ≈ 0.16), whereby the well-defined nature of the micelle was maintained upon extended incubation at 37 °C (36 h) and storage at 4 °C (1 month). Labeling of the micelles with Alexa Fluor 594 and Alexa Fluor 647, which form a Förster resonance energy transfer (FRET) pair, indicated the stability of loaded micelles upon dilution until the CMC. Finally, the cytotoxicity of the loaded micelles was investigated against three different cell lines: Medulloblastoma cell lines ONS-76 and DAOY as well as the glioblastoma cell line U87MG. While the panobinostat-loaded micelles displayed similar cytotoxicity compared to the pure drug in the cell lines, imatinib-loaded micelles were found to be more potent compared to the pristine drug, as significantly higher cytotoxicity was observed across all three cell lines.

Preparation and dynamic color-changing study of fluorescent polymer nanoparticles for individualized and customized anti-counterfeiting application

Preparing new fluorescent materials for individualized and customized anti-counterfeiting applications to meet needs from the rapid development of e-commerce is of great significance. This paper reports the preparation of dynamic color-changing fluorescent polymer nanoparticles (PNPs) by constructing a fluorescence resonance energy transfer (FRET) pair between aggregation-induced emission (AIE) structures and photochromic structures. At first, methyl methacrylate (MMA) was used as the main monomer and tetraphenylethylene (TPE, a typical AIE structure) modified methacrylate (TPE-MA) and photochromic spiropyran (SP) modified methacrylate (SP-MA) as minor monomers were copolymerized to obtain the ternary copolymer PMMA-TPE-SP. Then, two types of PNPs based on this terpolymer was prepared via the reprecipitation method, with and without the addition of an amphiphilic polymer as the surfactant. The photophysical study shows that the fluorescence color of PNPs dynamically changes from blue to light violet and finally to red under UV light irradiation, a process that can be reversed under visible light. The PNPs were alternately irradiated with UV light and visible light for 10 cycles, which proved their good photoswitching reproducibility. The PNPs prepared with addition of surfactant were found to have stronger fluorescence and better stability. Finally, the photochromic fluorescent inks were prepared based on these PNPs. Several anti-counterfeiting scenarios and modes were designed, exhibiting excellent photochromic behavior on cellulose paper, even after 120 days of long-term storage. With simple equipment, desirable anti-counterfeiting effects with dynamic fluorescence color changing was achieved. This study demonstrated a promising hard-to-imitate anti-counterfeiting encryption strategy, which can achieve multiple outputs with simple operation and can be personalized and customized as needed.

A general method for the development of multicolor biosensors with large dynamic ranges

Fluorescent biosensors enable the study of cell physiology with spatiotemporal resolution; yet, most biosensors suffer from relatively low dynamic ranges. Here, we introduce a family of designed Förster resonance energy transfer (FRET) pairs with near-quantitative FRET efficiencies based on the reversible interaction of fluorescent proteins with a fluorescently labeled HaloTag. These FRET pairs enabled the straightforward design of biosensors for calcium, ATP and NAD+ with unprecedented dynamic ranges. The color of each of these biosensors can be readily tuned by changing either the fluorescent protein or the synthetic fluorophore, which enables simultaneous monitoring of free NAD+ in different subcellular compartments following genotoxic stress. Minimal modifications of these biosensors furthermore allow their readout to be switched to fluorescence intensity, fluorescence lifetime or bioluminescence. These FRET pairs thus establish a new concept for the development of highly sensitive and tunable biosensors.

Enhanced resonance energy transfer in gold nanoparticles bifunctionalized by tryptophan and riboflavin and its application in fluorescence bioimaging

Gold nanoparticles were functionalized by amino acid tryptophan and vitamin riboflavin - a resonance energy transfer (RET) pair of biomolecules. The presence of the gold nanoparticles resulted in 65% increase in RET efficiency. Because of enhanced RET efficiency, the photobleaching dynamics of the fluorescent molecules at the surface of the nanoparticles is different from that of molecules in solution. The observed effect was used for detection of the functionalized nanoparticles within biological material rich with autofluorescent species. Synchrotron radiation deep-ultraviolet fluorescence microscopy is used to study the photobleaching dynamics of the fluorescence centers within human hepatocellular carcinoma Huh7.5.1 cells incubated with the nanoparticles. The fluorescent centers were classified according to their photobleaching dynamics, which enabled the discrimination of the cell areas where the accumulation of the nanoparticles takes place, even though the particles were smaller than the spatial resolution of the images.

Membrane Rigidity‐Tunable Fusogenic Nanosensor for High Throughput Detection of Fusion‐Competent Influenza A Virus

AbstractThe emergence of fatal viruses that pose continuous threats to global health has fueled the intense effort to develop direct, accurate, and high‐throughput virus detection platforms. Current diagnostic methods, including qPCR and rapid antigen tests, indicate how much of the virus is present, whether small fragments or whole viruses. However, these methods do not indicate the probability of the virus to be active, capable of interacting with host cells and initiating the infection cycle. Herein, a sialic acid‐presenting fusogenic liposome (sLipo–Chol) nanosensor with purposefully modulated membrane rigidity to rapidly detect the fusion‐competent influenza A virus (IAV) is developed. This nanosensor possesses virus‐specific features, including hemagglutinin (HA) binding and HA‐mediated membrane fusion. It is explored how the fusogenic capability of sLipo–Chol with different membrane rigidities impacts their sensing performance by integrating Förster resonance energy transfer (FRET) pairs into the bilayers. The addition of an intact virus led to instant FRET signal changes, thus enabling the direct detection of diverse IAV subtypes—even in avian fecal samples—within an hour at room temperature. Therefore, the sensing approach, with an understanding of the cellular pathogenesis of influenza viruses, will aid in developing bioinspired nanomaterials for evolution into nanosystems to detect infection‐competent viruses.

Rapid Detection of Ag(I) via Size-Induced Photoluminescence Quenching of Biocompatible Green-Emitting, l-Tryptophan-Scaffolded Copper Nanoclusters.

Atomically precise metal nanoclusters capped with small molecules like amino acids are highly favored due to their specific interactions and easy incorporation into biological systems. However, they are rarely explored due to the challenge of surface functionalization of nanoclusters with small molecules. Herein, we report the synthesis of a green-emitting (λex = 380 nm, λem = 500 nm), single-amino-acid (l-tryptophan)-scaffolded copper nanocluster (Trp-Cu NC) via a one-pot route under mild reaction conditions. The synthesized nanocluster can be used for the rapid detection of a heavy metal, silver (Ag(I)), in the nanomolar concentration range in real environmental and biological samples. The strong green photoluminescence intensity of the nanocluster quenched significantly upon the addition of Ag(I) due to the formation of bigger nanoparticles, thereby losing its energy quantization. A notable color change from light yellow to reddish-brown can also be observed in the presence of Ag(I), allowing its visual colorimetric detection. Portable paper strips fabricated with the Trp-Cu NC can be reliably used for on-site visual detection of Ag(I) in the micromolar concentration range. The Trp-Cu NC possesses excellent biocompatibility, making it a suitable nanoprobe for cell imaging; thus, it can act as an in vivo biomarker. The nanocluster showed a significant spectral overlap with anticancer drug doxorubicin and thus can be used as an effective fluorescence resonance energy transfer (FRET) pair. FRET results can reveal important information regarding the attachment of the drug to the nanocluster and hence its role as a potential drug carrier for targeted drug delivery within the human body.

Amphiphilic Near‐Infrared Fluorescent Molecular Probes: Optical Properties in Solution and in Surfactant Micelle Microenvironment

AbstractIn this work, the optical properties of two novel near‐infrared fluorescent tricarbocyanines with amphiphilic nature, CyN and CyS, were studied in solution and in micelles of representative surfactants (anionic, cationic and non‐ionic) by means of spectroscopic techniques. Interestingly, the spectral properties of the new fluorophores depend on the nature of the heteroatom at the meso position. Dependence on the solvent polarity and the viscosity were also observed in both cases. Furthermore, changes on photophysical properties occurred upon encapsulation of the dyes. The incorporation of the dye from the aqueous solution to the surfactant supramolecular assembly produced a drastic change in the fluorescence emission from quenched state to fluorescence enhancement, rendering an OFF‐ON response to the exchange of environment. In addition, a significant protective effect against photobleaching was determined when the dyes were included in Pluronic® micelles (F127) resulting in a promising system to be used as a fluorescent nanosized container. The novel amphiphilic NIR fluorophores also interact as a Förster resonance energy transfer (FRET) pair and the process was studied within micelles loaded with both dyes.

Förster Resonance Energy Transfer Nanobullet for Photoacoustic Imaging and Amplified Photothermal-Photodynamic Therapy of Cancer.

Synergistic photodynamic and photothermal therapy (PDT-PTT) has emerged as an appealing effective antitumor approach. However, clinical utilization of PDT-PTT is plagued by aggregation-caused photobleaching, sequential double irradiations, unsatisfying balance between single oxygen (1 O2 ) quantum yield and photothermal conversion efficiency. Here, an anchored tumor-homing cell-penetrating peptide (PEGA-pVEC) and PANI-ES/HMME loaded FRET nanobullet (AHP-P) are reported. Within nanobullet, HMME (donor) and PANI-ES (acceptor) spontaneously form a förster resonance energy transfer (FRET) pair. Upon 660nm laser irradiation, HMME convert near-infrared fluorescence (NIRF) to PANI, thus produce FRET-amplified photoacoustic imaging guided PTT. In addition, AHP-P with pH-sensitivity can gradually release HMME within acidic tumor environment, boosts the 1 O2 regeneration alongside with highly efficient photothermal conversion for photoinduced cancer PTT-PDT. Furthermore, the AHP-P nanobullet can home in on the tumor site and penetrate into cytoplasm through PEGA-pVEC, inducing remarkable tumor regression with an ≈80% tumor volume reduction and decreased skin phototoxicity in vivo during FRET-amplified PTT-PDT.

Mechano-fluorescence actuation in single synaptic vesicles with a DNA framework nanomachine.

Biomimetic machines that can convert mechanical actuation to adaptive coloration in a manner analogous to cephalopods have found widespread applications at various length scales. At the nanoscale, a transmutable nanomachine with adaptive colors that can sense and mediate cellular or intracellular interactions is highly desirable. Here, we report the design of a DNA framework nanomachine (DFN) that can autonomously change shape in response to pH variations in single synaptic vesicles, which, in turn, displays adaptive fluorescent colors with a mechano-fluorescence actuation mechanism. To construct a DFN, we used a tetrahedral DNA nanostructure as the framework to incorporate an embedded pH-responsive, i-motif sequence tagged with a Förster resonance energy transfer pair and an affinity cholesterol moiety targeting vesicular membranes. We found that endocytosed DFNs are individually trapped in single endocytic vesicles in living synaptic cells due to the size-exclusion effect. The adaptive fluorescence coloration of DFNs enabled single-vesicle quantification of resting pH values in a processive manner, allowing long-term tracking of the exocytosis and fusion dynamics in intracellular processes and cell-cell communications.

Peptide Array-Based In Situ Fluorescence Assay for Profiling Multiple Matrix Metalloproteinase Activities.

Peptide array-based in situ fluorescence assay is a reliable and efficient technique for high-throughput profiling and localization of enzyme activity. Here, peptide array is fabricated by spotting five specific MMPs (MMP-2, MMP-3, MMP-7, MMP-9, and MMP-14) peptide substrates containing FAM/Dabcyl fluorescent resonance energy transfer (FRET) pair on the surface of cell monolayers or tissue sections. MMP activities are determined in situ by the fluorescence intensity of stained cells/tissues due to the cellular internalization of hydrolyzed peptide fragments with FAM moieties. Identification of MMP expression patterns of cells, highly sensitive determination of MMP activities in cell monolayer (as low as hundreds of cells per square centimeter), and evaluation of inhibition potencies of six compounds toward five MMPs are achieved by this method. Five MMP activities in the localized parts of 32 thyroid tissues are also well profiled without separation or extraction procedures.

PDT-active upconversion nanoheaters for targeted imaging guided combinatorial cancer phototherapies with low-power single NIR excitation

A versatile nanoformulation is designed by anchoring human transferrin protein (Tf) on fluoromagnetic upconverting nanoheaters, NaGdF4:Yb,Er (UCNP), loaded with Rose Bengal (RB), for multimodal imaging guided synergistic photothermal (PTT) and photodynamic therapy (PDT) at the targeted tumor site. The NIR excitation of the UCNP-RB Forster Resonance Energy Transfer (FRET) pair results in the reactive oxygen species (ROS) generation for PDT, whereas the non-radiative transitions in Er result in the heat required for PTT. The intravenously injected theranostic agent (UCNP@Tf-RB) enabled; (1) combinatorial PTT and PDT of 4T1 tumors with minimal systemic toxicity, (2) dual targeted (passive and active) tumor accumulation, (3) dual-modal imaging (MRI/photothermal), and, (4) excellent stability and biocompatibility. The in vitro therapy data corroborates the MRI findings that Tf conjugation resulted in actively targeted tumor accumulation via over-expressed transferrin receptors (TfR) on 4T1 cells. Real-time photothermal imaging enabled visualization of the tumor while receiving the therapy. The UCNP@Tf-RB, for synergistic PTT-PDT, and UCNP@Tf, for PTT only, caused rapid suppression of tumor with a tumor-growth inhibition index (TGII) of ~0.91, and 0.79, respectively. Histopathological examination demonstrated minimal damage to non-targeted tissues and caused significant damage to the tumor. This theranostic methodology enhances anti-cancer therapeutic efficiency, and announces the potential for pre-clinical cancer therapy.

Radioluminescent Photonic Bandgap Hydrogels: Mechanochromic Tunable Emissions.

Fully organic, radioluminescent crystalline colloidal arrays (CCAs) with covalently incorporated emitters were synthesized by using up to three organic fluorophores that were Förster resonance energy transfer (FRET) pairs with each other. The emitters were covalently incorporated into monodisperse poly(styrene-co-propargyl acrylate) nanoparticles in various combinations, resulting in blue-, green-, and red-emitting CCAs when excited with an X-ray source. The negatively charged surfaces of the monodisperse nanoparticles caused self-assembly into a crystal-like structure, which resulted in a partial photonic bandgap (i.e., rejection wavelength) within the near-visible and visible light spectrum. When the rejection wavelength of the CCA overlapped its radioluminescence, the spontaneous emission was inhibited and the emission intensity decreased. A poly(ethylene glycol) methacrylate-based hydrogel network was used to encapsulate the CCAs and stabilize their crystal-like structure. Within the hydrogel, coupling the photonic bandgap with the radioluminescence of the CCA films led to robust optical systems with tunable emissions. These fully organic, hydrogel-stabilized, radioluminescent CCAs possess mechanochromic tunable optical characteristics with future applications as potentially less toxic X-ray bioimaging materials.

Conformational changes of exogenous deposited fibronectin on decellularized aortic conduits after implantation

Fibronectin is an abundant and ubiquitous extracellular matrix component that regulates a wide range of cell functions by directly interacting with cell surface integrin receptors. In a prior study, we observed that fibronectin can accelerate the autologous cellular repopulation of implanted decellularized grafts, enhancing their adhesion capacity and biocompatibility, through fibronectin fibrillogenesis. The supportive effect of deposited fibronectin was shown to be dissimilar in different regions of implanted decellularized grafts. This led us to question whether there is any difference in conformation of fibronectin deposited on the luminal and adventitial surfaces of decellularized aortic conduits after implantation. In this study, we doubly labelled fibronectin with a fluorescent resonance energy transfer (FRET) pair and use this to analyse differences in conformational changes of adsorbed fibronectin at different sites of the decellularized aortic conduits.

Making the most of DNA melt curves: data collapse with affine transformations and consequences of experimental design

Nucleic acid melt data is ubiquitous, whether obtained by ultraviolet-visible spectroscopy (UV-vis), fluorescence from Förster Resonance Energy Transfer (FRET) pairs, intercalating dyes, or other reporters, and, in the context of quantitative Polymerase Chain Reaction (qPCR) experiments and High-Resolution Melting (HRM) experiments, plentiful. While van’t Hoff analysis of nucleic acid melting data is considered to be straightforward, considerable care is needed to ensure that it yields precise and accurate thermodynamic parameters which are consistent with other measurements and useful for prediction.

FRET measurement of cytochrome bc1 and reaction centre complex proximity in live Rhodobacter sphaeroides cells

In the model purple phototrophic bacterium Rhodobacter (Rba.) sphaeroides, solar energy is converted via coupled electron and proton transfer reactions within the intracytoplasmic membranes (ICMs), infoldings of the cytoplasmic membrane that form spherical ‘chromatophore’ vesicles. These bacterial ‘organelles’ are ideal model systems for studying how the organisation of the photosynthetic complexes therein shape membrane architecture. In Rba. sphaeroides, light-harvesting 2 (LH2) complexes transfer absorbed excitation energy to dimeric reaction centre (RC)-LH1-PufX complexes. The PufX polypeptide creates a channel that allows the lipid soluble electron carrier quinol, produced by RC photochemistry, to diffuse to the cytochrome bc1 complex, where quinols are oxidised to quinones, with the liberated protons used to generate a transmembrane proton gradient and the electrons returned to the RC via cytochrome c2. Proximity between cytochrome bc1 and RC-LH1-PufX minimises quinone/quinol/cytochrome c2 diffusion distances within this protein-crowded membrane, however this distance has not yet been measured. Here, we tag the RC and cytochrome bc1 with yellow or cyan fluorescent proteins (YFP/CFP) and record the lifetimes of YFP/CFP Förster resonance energy transfer (FRET) pairs in whole cells. FRET analysis shows that that these complexes lie on average within 6 nm of each other. Complementary high-resolution atomic force microscopy (AFM) of intact, purified chromatophores verifies the close association of cytochrome bc1 complexes with RC-LH1-PufX dimers. Our results provide a structural basis for the close kinetic coupling between RC-LH1-PufX and cytochrome bc1 observed by spectroscopy, and explain how quinols/quinones and cytochrome c2 shuttle on a millisecond timescale between these complexes, sustaining efficient photosynthetic electron flow.

Organic Light‐Emitting Diode Based Fluorescence Sensing System for DNA Detection

AbstractConventional fluorescence sensing equipment for disease detection is expensive and bulky, restricting access of patients to accurate diagnosis. Organic light‐emitting diodes (OLEDs) have the potential to enable compact fluorescence sensing compatible with point‐of‐care (POC) testing. However, the limited brightness and broad emission spectra of OLEDs can be a challenge for achieving good sensitivity. Here, co‐host microcavity OLEDs with narrowed spectra, high conductivity, and high brightness are developed for fluorescence sensing. The OLEDs are driven in pulsed mode for achieving higher brightness and stable light output. To sense the presence of single‐stranded DNA (ssDNA), two complementary ssDNA labelled with Cyanine dyes – Cy3 (ssDNA‐Cy3) or Cy5 (ssDNA‐Cy5) are used to form a Förster resonance energy transfer (FRET) pair when the DNA hybridized. The dye‐labelled DNA is then excited by the OLED at the Cy3 excitation wavelength, and the fluorescence is detected at the emission wavelength of Cy5. As a result, the device shows a very high sensitivity that can detect as low as 1 × 10−9 m of ssDNA‐Cy5 in fetal bovine serum (FBS). This work shows a simple approach to highly sensitive fluorescence sensing with OLED light sources that is promising for use in POC diagnostics.

Super-Assembled Periodic Mesoporous Organosilica Frameworks for Real-Time Hypoxia-Triggered Drug Release and Monitoring.

Hypoxia, induced by inadequate oxygen supply, is a key indication of various major illnesses, which necessitates the need to develop new nanoprobes capable of sensing hypoxia environments for the targeted system monitoring and drug delivery. Herein, we report a hypoxia-responsive, periodic mesoporous organosilica (PMO) nanocarrier for repairing hypoxia damage. β-cyclodextrin (β-CD) capped azobenzene functionalization on the PMO surface could be effectively cleaved by azoreductase under a hypoxia environment. Moreover, the nanosystem is equipped with fluorescence resonance energy transfer (FRET) pair (tetrastyrene derivative (TPE) covalently attached to the PMO framework as the donor and Rhodamine B (RhB) in the mesopores as the receptor) for intracellular visualization and tracking of drug release in real-time. The design of intelligent nanocarriers capable of simultaneous reporting and treating of hypoxia conditions highlights a great potential in the biomedical domain.