Dual-modality Probe Research Articles

Fluoropolymer-Gadolinium(III) Hybrids for Photoactivatable Dual-Mode T1-Weighted 1H MRI and 19F MRI Contrast Agents.

Multimodal imaging probes play a crucial role in overcoming the limitations associated with single-mode imaging for clinical medical diagnosis. This study focuses on the development of a photoresponsive fluorine-containing water-soluble polymer (PF) through RAFT polymerization. Subsequently, a polymer-gadolinium(III) hybrid (PF-Gd) dual-modal probe capable of T1-weighted 1H MRI and 19F MRI was synthesized via postmodification of PF with a Gd-DOTA derivative. Under physiological conditions (pH = 7.4), the hybrids exhibit UV-activated 19F NMR/MRI and enhanced 1H MRI. The inclusion of Gd3+ facilitates the acceleration of water molecule T1 relaxation, leading to high-intensity 1H MRI contrast. Leveraging the paramagnetic relaxation enhancement (PRE) effect between fluorine atoms and Gd3+, the restoration of Gd3+-accelerated 19F T2 relaxation enables precise photoactivation of 19F MRI signals, transitioning from the "OFF" to the "ON" state. This study provides an important reference for the development of hybrid systems that function as real-time diagnostic tools and offers controlled activation for multimodal imaging probes.

Bimetallic metal organic frameworks-based nanozyme for colorimetry/fluorescence dual-mode uremic toxin detection

Chronic kidney disease (CKD) can greatly increase the mortality risk for clinical patients, with a major challenge being the accurate detection of uremic toxins. Metal organic frameworks (MOFs)-based colorimetry/fluorescence dual-mode probe have shown great potential for bio-detection. However, one of the challenges for MOFs-based nanoprobe is to effectively improve the activity of mimetic enzymes for colorimetric assays while maintaining their outstanding luminescence response for fluorescence sensing. Here, we developed a novel bimetallic Zr/Ce-MOFs, which possesses high catalytic activity and excellent luminescence properties for dual-mode sensing of uremic toxins. The co-existence of Ce and Zr in the MOFs improves the catalytic activity for colorimetric detection of phosphate ions (one type of uremic toxins) and the strong coordinate covalent interaction between phosphate ions and Zr/Ce leads to a significant fluorescence enhancement of the MOFs. Furthermore, we achieved high sensitivity phosphate ions analysis in serum samples. This strategy provides an easy and reliable platform for the detection of uremic toxins in complicated physiological environment.

Controllable fabrication of high-quantum-yield bimetallic gold/silver nanoclusters as multivariate sensing probe for Hg2+, H2O2, and glutathione based on AIE and peroxidase mimicking activity

The grave threat posed by heavy metals and food hazards has increased the urgency of rapid and precise detection for food security and human health. Efficient multivariate sensing probes are imperatively required for sensing heavy metals and tumor markers, which are still facing great challenge in terms of multi-functional integration. Here, bimetallic gold-silver nanoclusters (NG-AuAgNCs) were developed with unique aggregation-induced-emission (AIE) property and peroxidase (POD) mimicking activity towards the efficient multivariate sensing via optimization of the precursors. The NG-AuAgNCs emitted at 614 nm and enable AIE feature with lifetime of 12.61 μs and high quantum yield of 40.5%. Possessing AIE and POD activity, the NG-AuAgNCs show great potential as fluorimetric and colorimetric dual-mode probe for multivariate sensing Hg2+, H2O2 and GSH, with good recoveries in real samples. The NG-AuAgNCs paper sensors further integrating with smartphone, achieved portable detection of Hg2+ with limit of detection (LOD) of 19 nM, while the colorimetric-mode presented consecutive response to H2O2 and GSH via a reversible oxidase tetramethylbenzidine process with LODs of 7.02 and 0.45 μM, respectively. This work not only demonstrates a multivariate probe for environment and human health, but also provides valuable insights for the function integration of the nanocluster via synthetic manipulation.

In vivo targeted-imaging of mitochondrial acidification in an aristolochic acid I-induced nephrotoxicity mouse model by a fluorescent/photoacoustic bimodal probe

Aristolochic acid I (AAI), a natural compound in aristolochia type Chinese medicinal herb, is generally acknowledged to have nephrotoxicity, which may be associated with mitophagy. Mitophagy is a cellular process with important functions that drive AAI-induced renal injury. Mitochondrial pH is currently measured by fluorescent probes in cell culture, but existing probes do not allow for in situ imaging of AAI-induced mitophagy in vivo. We developed a ratiometric fluorescent/PA dual-modal probe with a silicon rhodamine fluorophore and a pH-sensitive hemicyanine dye covalently linked via a short chain to obtain a FRET type probe. The probe was used to measure AAI-mediated mitochondrial acidification in live cells and in vivo. The Förster resonance energy transfer (FRET)-mediated ratiometric and bimodal method can efficiently eliminate signal variability associated with the commonly used one-emission and single detection mode by ratiometric two channels of the donor and acceptor. The probe has good water-solubility and low molecular weight with two positively charged, facilitating its precise targeting into renal mitochondria, where the fluorescent/PA changes in response to mitochondrial acidification, enabling dynamic and semi-quantitative mapping of subtle changes in mitochondrial pH in AAI-induced nephrotoxicity mouse model for the first time. Also, the joint use of L-carnitine could mitigate the mitophagy in AAI-induced nephrotoxicity.

Fluorescence and Circular Dichroism Dual-Mode Probe for Chiral Recognition of Tyrosine and Its Applications in Bioimaging.

Chiral amino acids (AAs) are essential in metabolism and understanding physiological processes, and they could be used as biomarkers for the diagnosis of different diseases. In this study, chiral Cdots@Van were prepared by postmodifying an achiral Cdots core with vancomycin for recognizing and determining the enantiomeric excess (ee) of tyrosine (Tyr) enantiomers. The fluorescence response of Cdots@Van is based on an "on-off" strategy, with different quenching percentages for d- and l-tyrosine. Interestingly, the circular dichroism (CD) spectrum of Cdots@Van responded to only one form of Tyr enantiomer, specifically d-Tyr, and remained nearly unchanged upon the addition of l-Tyr. Quantum mechanical (QM) calculations were in excellent agreement with the experimental results, confirming the stronger binding affinity of Cdots@Van for d-Tyr compared to l-Tyr. We further investigated the chiral recognition ability of the interconnected vancomycin particles, which was synthesized using the EDC/NHS coupling reaction between vancomycin molecules without a Cdots core. Surprisingly, unlike free vancomycin molecules, interconnected vancomycin displayed an enantiomeric recognition ability by CD spectroscopy, similar to what was observed for Cdots@Van. Crucially, this chiral probe has been successfully utilized for cell imaging applications.

High-fidelity imaging of drug-induced acute gastritis by using a fluorescent and photoacoustic dual-modal probe with good stability in stomach acid

In consideration of deep tissue imaging and signal fidelity, fluorescent-photoacoustic (PA) dual-modal probes are much more desirable. However, dual-modal imaging of gastritis using molecular probes remains a challenge due to the harsh gastric acid environment in the stomach. Based on the positive correlation between gastritis and cell viscosity, stomach acid-stable and viscosity-activated probes could potentially diagnose gastritis. As a proof of concept, herein, a fluorescent and photoacoustic dual-modal probe (named WSP-1) is revealed for the imaging of drug-induced acute gastritis in vivo. WSP-1 exhibits viscosity-dependent fluorescence emission and photoacoustic signals. A rotatable C–C single bond is incorporated into the D-π-A structure of WSP-1, which could facilitate the formation of the twisted intramolecular charge transfer (TICT) state in a low-viscosity environment (weak fluorescence/PA signal) and the intramolecular charge transfer (ICT) state in a high-viscosity environment (strong fluorescence/PA signal). WSP-1 has demonstrated the capability to target mitochondria and can be utilized to monitor the viscosity enhancement of cells during inflammation. Most importantly, WSP-1 exhibits good optical and structural stability in gastric acid. By leveraging these desirable features of WSP-1, we have achieved fluorescent and 3D photoacoustic in situ imaging of drug-induced acute gastritis following oral administration of WSP-1.

A turn-on anthraquinone-derived colorimetric and fluorometric dual-mode probe for highly selective Hg2+ determination and bioimaging in living organisms

Mercury ion (Hg2+) is considered a harmful neurotoxin, and real-time monitoring of Hg2+ concentrations in environmental and biological samples is critical. Fluorescent probes are a rapidly emerging visualization tool owing to their simple design and good selectivity. Herein, a novel fluorescence (FL) probe 2-(4-((6-((quinolin-8-yloxy)methyl)pyridin-2-yl)methyl)piperazin-1-yl)anthracene-9,10-dione (QPPA) is designed using piperazine as a linker between the anthraquinone group, which serves as a fluorophore, and N4O as the Hg2+ ligand. The probe exhibits FL “turn-on” sensing of Hg2+ because the complex inhibits the photo-induced electron transfer (PET) process. Moreover, QPPA can overcome the invasion by other possible cations, resulting in a clear color change from orange to colorless with the addition Hg2+. The chelation of QPPA with Hg2+ in a 1:1 ratio. Subsequently, the theoretically determined binding sites of the ligand to Hg2+ are validated through 1H NMR titration. The in situQPPA–Hg2+ complex can be subjected to Hg2+ extraction following the introduction of S2− owing to its robust binding capacity. The exceptional limit of detection values for Hg2+ and S2− are obtained as 63.0 and 79.1 nM (S/N = 3), respectively. Moreover, QPPA can display bright red FL in the presence of Hg2+ in different biological specimens such as HeLa cells, zebrafish, onion root tip tissues, and water flea Daphnia carinata, further providing an effective strategy for environmental monitoring and bioimaging of Hg2+ in living organisms.

Preclinical Evaluation of a PSMA Aptamer-Based Bifunctional PET and Fluorescent Probe.

Prostate cancer is the most prevalent malignant tumor affecting male individuals worldwide. The accurate early detection of prostate cancer is crucial to preventing unnecessary diagnosis and subsequent excessive treatment. Prostate-specific membrane antigen (PSMA) has emerged as a promising biomarker for the diagnosis of prostate cancer. In this study, a dual-modality imaging probe utilizing aptamer technology was developed for positron emission tomography/near-infrared fluorescence (PET/NIRF) imaging, and the specificity and sensitivity of the probe toward PSMA were evaluated both in vitro and in vivo. The probe precursor NOTA-PSMA-Cy5 was synthesized via automated solid-phase oligonucleotide synthesis. Subsequently, the PET/NIRF dual-modality probe [68Ga]Ga-NOTA-PSMA-Cy5 was successfully prepared and exhibited favorable fluorescence properties and stability in vitro. The binding specificity of [68Ga]Ga-NOTA-PSMA-Cy5 to PSMA was assessed through flow cytometry, fluorescence imaging, and cellular uptake experiments in LNCaP cells and PC-3 cells. In vivo PET/NIRF imaging studies demonstrated the sensitive and specific binding of [68Ga]Ga-NOTA-PSMA-Cy5 to PSMA. Overall, the PET/NIRF dual-modality probe [68Ga]Ga-NOTA-PSMA-Cy5 shows promise for the diagnosis of prostate cancer and for the fluorescence-guided identification of PSMA-positive cancer lesions during surgical procedures.

Fluorescence and photoacoustic (FL/PA) dual-modal probe: Responsive to reactive oxygen species (ROS) for atherosclerotic plaque imaging

Accurate and early detection of atherosclerosis (AS) is imperative for their effective treatment. However, fluorescence probes for efficient diagnosis of AS often encounter insufficient deep tissue penetration, which hinders the reliable assessment of plaque vulnerability. In this work, a reactive oxygen species (ROS) activated near-infrared (NIR) fluorescence and photoacoustic (FL/PA) dual model probe TPA-QO-B is developed by conjugating two chromophores (TPA-QI and O–OH) and ROS-specific group phenylboronic acid ester. The incorporation of ROS-specific group not only induces blue shift in absorbance, but also inhibits the ICT process of TPA-QO-OH, resulting an ignorable initial FL/PA signal. ROS triggers the convertion of TPA-QO-B to TPA-QO-OH, resulting in the concurrent amplification of FL/PA signal. The exceptional selectivity of TPA-QO-B towards ROS makes it effectively distinguish AS mice from the healthy. The NIR emission can achieve a tissue penetration imaging depth of 0.3 cm. Moreover, its PA775 signal possesses the capability to penetrate tissues up to a thickness of 0.8 cm, ensuring deep in vivo imaging of AS model mice in early stage. The ROS-triggered FL/PA dual signal amplification strategy improves the accuracy and addresses the deep tissue penetration problem simultaneously, providing a promising tool for in vivo tracking biomarkers in life science and preclinical applications.

Activatable near-infrared fluorescent/photoacoustic probe for rapid identification of β-lactam-resistant bacteria

Early identification of antimicrobial-resistant (AMR) bacteria is of great significance to guide the use of drugs and reduce the risk of AMR infection. One of the main reasons for AMR resistance to antibiotic therapy is the production of β-lactamase that decomposes antibiotics. Accurate and rapid detection of β-lactamase is crucial o deal with bacterial resistance. Herein, a dual-modal probe with near-infrared fluorescence (NIRF) and photoacoustics (PA) to rapid detection of β-lactamases (TEM-1) and β-lactam-resistant bacteria (BLRB) was ingeniously designed and developed. The NIRF and PA signal of CyPA-L can be fully turned on after 7 min of incubation of CyPA-L with TEM-1, and also showed high selectivity for TEM-1 among multiple interfering analytes. Notably, a portable PA device with immediate detection (POCT) capability was developed to detect β-lactamase and BLRB through PA signaling of CyPA-L. The design of probe and facile device will provide a new way for the diagnosis and symptomatic treatment of early drug-resistant bacteria.

Rare-earth-free up and down-conversion dual-emission carbon dots for Cu2+ sensing

In this work, up- and down-conversion dual-emission CDs without rare-earth (UD D-CDs) were synthesized using RhB and 1,4-Diaminoanthraquinone as precursors. The synthesized UD D-CDs exhibited dual emissions at 496 and 580 nm under 260 and 865 nm excitation, respectively. The fluorescence emission mechanism, including contributions from carbon nuclei, surface states, molecular states, and internal defect states, was discussed through the separation and purification of UD D-CDs. Based on the interaction between UD D-CDs and copper ions (Cu2+), a dual-mode ratio fluorescence probe was developed to detect and quantify Cu2+. The up-conversion ratio fluorescent probe shows a linear range of 0.0500–15.0 μM, with a detection limit as low as 2.76 nM. This method has been successfully applied to detecting Cu2+ in human serum and has potential applications in biochemical analysis and biological imaging. The successful preparation of up-conversion fluorescent carbon dots without rare earth elements and the ability to perform low-damage detection in high-background biological samples provide a new approach to constructing non-rare earth up-conversion probes.

Rational design of a dual-mode fluorescent probe for portable detection of pyriproxyfen in the environment and food

The development of a fluorescent probe for pyriproxyfen (PPF) is crucial due to its potential threat to human health. However, the chemical inertness and low solubility of PPF present significant challenges for the detection of PPF in aqueous solutions using fluorescent probes. Herein, we have originally proposed a complex based on 2-(4-(dimethylamino)phenyl)−3-hydroxy-6,7-dimethoxy-4 H-chromen-4-one (HOF) and serum albumin (SA) as a dual-mode fluorescent probe, HOF@SA. This probe utilizes an indicator displacement assay (IDA) to release the dye HOF from the probe at low PPF concentrations (< 10 µM) and embeds the free dye HOF into the micelle of PPF at high concentrations (> 10 µM). This results in dual-mode fluorescent response characteristics for PPF: a turn-off response at low concentrations and a ratiometric response at high concentrations. An investigation of sensing behavior of HOF@SA for PPF detection exhibits rapid response (< 60 s), high sensitivity (LOD ∼4.7 ppb), high selectivity, and excellent visual detection capability (from cyan to yellow). Moreover, with the aid of a portable device, this method enables to analyze PPF in environmental and food samples. These results promote the advancement of a fluorescent probe approach for PPF analysis in environment and food.

A novel fluorescent and photothermal probe based on nanozyme-mediated cascade reaction for detecting organophosphorus pesticide residues

In this study, a nanozyme (ZIF-Co-Cys) with high oxidase-like catalytic activity was prepared, and a ratiometric fluorescent/photothermal dual-mode probe was constructed for organophosphorus pesticides (OPs) detection based on the competitive effect of ZIF-Co-Cys and the enzymatic reaction product of acid phosphatase (ACP) on o-phenylenediamine and the inhibition effect of OPs on ACP activity. Using dimethyl dichloroviny phosphate (DDVP) as the model, both the fluorescence intensity ratio and the temperature change of the probe solution exhibited an excellent correlation with OPs concentration. The detection limits were 1.64 ng/mL and 0.084 ng/mL, respectively. Additionally, the detection of DDVP residues in real samples verified the outstanding anti-interference and accuracy of the probe. This work not only provided a complementary dual-mode method for the accurate and rapid detection of OPs residues in complex samples, but also supplied a new insight into the design of a multi-mode sensing platform based on the cascade reaction of nanozyme.

High-fidelity imaging of fatty liver in vivo with a fluorescent and photoacoustic dual-mode probe by taking advantage of the liver “first pass effect”

Intravenous injection is the most commonly used method for probe delivery in bio-imaging. While, the oral administration method might be more efficient than intravenous injection for fatty liver imaging due to the existence of liver “first pass effect”. However, this fundamental question has not yet been clarified. Meanwhile, in consideration of the deep tissue imaging and signal fidelity, fluorescent-photoacoustic (PA) dual-mode probes are much more desirable. Herein, an orally administrable dual-mode probe WSP-3 is revealed as a tool for imaging of the fatty liver disease and providing an insight into the above scientific question. A D-π-A system is linked with a rotatable CC single bond in WSP-3, which facilitates the formation of a TICT state in low viscosity environments and an ICT state in high viscosity environments. Base on the positive correlation between fatty liver disease and cell viscosity, dual-mode imaging of non-alcoholic fatty liver disease (NAFLD) and drug-induced fatty liver disease (DIFLD) in vivo was accomplished. Moreover, control experiments indicated that the oral administration method demonstrated higher efficiency in fatty liver disease imaging, providing a fundamental reference for further probe-based fatty liver imaging.

A Wearable Dual-Mode Probe for Image-Guided Closed-Loop Ultrasound Neuromodulation.

Low-intensity focused ultrasound (FUS) is an emerging non-invasive and spatially/temporally precise method for modulating the firing rates and patterns of peripheral nerves. This paper describes an image-guided platform for chronic and patient-specific FUS neuromodulation. The system uses custom wearable probes containing separate ultrasound imaging and modulation transducer arrays realized using piezoelectric transducers assembled on a flexible printed circuit board (PCB). Dual-mode probes operating around 4 MHz (imaging) and 1.3 MHz (modulation) were fabricated and tested on tissue phantoms. The resulting B-mode images were analyzed using a template-matching algorithm to estimate the location of the target nerve and then direct the modulation beam toward the target. The ultrasound transmit voltage used to excite the modulation array was optimized in real-time by automatically regulating functional feedback signals (the average rates of emulated muscle twitches detected by an on-board motion sensor) through a proportional and integral (PI) controller, thus providing robustness to inter-subject variability and probe positioning errors. The proposed closed-loop neuromodulation paradigm was experimentally demonstrated in vitro using an active tissue phantom that integrates models of the posterior tibial nerve and nearby blood vessels together with embedded sensors and actuators.

Construction of multifunctional core-shell structure nanoprobe integrating NIR dual-mode optical thermometer and nuclear magnetic imaging

Precise temperature regulation and imaging are critical for in vivo photothermal treatment. The goal of this work is to realize non-contact temperature measurement and imaging in biological tissues by building a multifunctional nano-platform using optical temperature probes and imaging for biological diagnosis and therapy. Here, NIR dual-mode temperature probes of NaYF4: Nd/Yb/Gd@NaNdF4 were designed according to the LIR-I of 4F3/2 → 4I9/2/2F5/2 → 2F7/2 levels in 850 – 1025 nm and LIR-II of 2F5/2 → 2F7/2/4F3/2 → 4I11/2 in 950 – 1100 nm from the emissions of Nd3+/Yb3+. Transmission electron microscope (TEM) and X-ray diffraction (XRD) data of the sample demonstrate its superior dispersibility and accurate structure. The integrated intensity ratio centered around various luminescence intensity peaks is used as a temperature sensor since luminescence intensity is temperature-dependent. Additionally, the temperature sensing capabilities of two ratio temperature sensors were examined under 808 nm laser excitation, utilizing LIR (luminescence intensity ratio) technology. Furthermore, the MRI test indicated that the sample exhibited good imaging capacity when Gd3+ was added. These results assure the auspicious application prospects of the samples in temperature sensing, photothermal therapy, and biomedical imaging.

Pretheranostic agents with extraordinaryNIRF/photoacoustic imaging performanceand photothermal oncotherapy efficacy

Cervical cancer, the most common gynecological malignancy, significantly and adversely affects women's physical health and well-being. Traditional surgical interventions and chemotherapy, while potentially effective, often entail serious side effects that have led to an urgent need for novel therapeutic methods. Photothermal therapy (PTT) has emerged as a promising approach due to its ability to minimize damage to healthy tissue. Connecting a biothiol detection group to PTT-sensitive molecules can improve tumor targeting and further minimize potential side effects. In this study, we developed a near-infrared fluorescence (NIRF)/photoacoustic (PA) dual-mode probe, S-NBD, which demonstrated robust PTT performance. This innovative probe is capable of activating NIRF/PA signals to enable the detection of biothiols with high emission wavelength (838 nm) and large Stokes shift (178 nm), allowing for in vivo monitoring of cancer cells. Additionally, the probe achieved an outstanding photothermal conversion efficiency of 67.1%. The application of laser irradiation (660 nm, 1.0 W/cm2, 5 min) was able to achieve complete tumor ablation without recurrence. In summary, this seminal study presents a pioneering NIRF/PA dual-mode dicyanoisophorone-based probe for biothiol imaging, incorporating features from PTT for the first time. This pioneering approach achieves the dual objectives of improving tumor diagnosis and treatment.

Carboxylesterase Activatable Molecular Probe for Personalized Treatment Guidance by Analyte-Induced Molecular Transformation.

Accurate visualization of tumor microenvironment is of great significance for personalized medicine. Here, we develop a near-infrared (NIR) fluorescence/photoacoustic (FL/PA) dual-mode molecular probe (denoted as NIR-CE) for distinguishing tumors based on carboxylesterase (CE) level by an analyte-induced molecular transformation (AIMT) strategy. The recognition moiety for CE activity is the acetyl unit of NIR-CE, generating the pre-product, NIR-CE-OH, which undergoes spontaneous hydrogen atom exchange between the nitrogen atoms in the indole group and the phenol hydroxyl group, eventually transforming into NIR-CE-H. In cellular experiments and in vivo blind studies, the human hepatoma cells and tumors with high level of CE were successfully distinguished by both NIR FL and PA imaging. Our findings provide a new molecular imaging strategy for personalized treatment guidance.

Tuning Tumor Targeting and Ratiometric Photoacoustic Imaging by Fine-Tuning Torsion Angle for Colorectal Liver Metastasis Diagnosis.

Photoacoustic (PA) tomography is an emerging biomedical imaging technology for precision cancer medicine. Conventional small-molecule PA probes usually exhibit a single PA signal and poor tumor targeting that lack the imaging reliability. Here, we introduce a series of cyanine/hemicyanine interconversion dyes (denoted Cy-HCy) for PA/fluorescent dual-mode probe development that features optimized ratiometric PA imaging and tunable tumor-targeting ability for precise diagnosis and resection of colorectal cancer (CRC). Importantly, Cy-HCy can be presented in cyanine (inherent tumor targeting and long NIR PA wavelength) and hemicyanine (poor tumor targeting and short NIR PA wavelength) by fine-tuning torsion angle and the ingenious transformation between cyanine and hemicyanine through regulation optically tunable group endows the NIR ratiometric PA and tunable tumor-targeting properties. To demonstrate the applicability of Cy-HCy dyes, we designed the first small-molecule tumor-targeting and NIR ratiometric PA probe Cy-HCy-H2S for precise CRC liver metastasis diagnosis, activated by H2S (a CRC biomarker). Using this probe, we not only visualized the subcutaneous tumor and liver metastatic cancers in CRC mouse models but also realized PA and fluorescence image-guided tumor excision. We expect that Cy-HCy will be generalized for creating a wide variety of inherently tumor-targeting NIR ratiometric PA probes in oncological research and practice.

A firm-push-to-open and light-push-to-lock strategy for a general chemical platform to develop activatable dual-modality NIR-II probes.

Activatable near-infrared (NIR) imaging in the NIR-II range is crucial for deep tissue bioanalyte tracking. However, designing such probes remains challenging due to the limited availability of general chemical strategies. Here, we introduced a foundational platform for activatable probes, using analyte-triggered smart modulation of the π-conjugation system of a NIR-II-emitting rhodamine hybrid. By tuning the nucleophilicity of the ortho-carboxy moiety, we achieved an electronic effect termed "firm-push-to-open and light-push-to-lock," which enables complete spirocyclization of the probe before sensing and allows for efficient zwitterion formation when the light-pushing aniline carbamate trigger is transformed into a firm-pushing aniline. This platform produces dual-modality NIR-II imaging probes with ~50-fold fluorogenic and activatable photoacoustic signals in live mice, surpassing reported probes with generally below 10-fold activatable signals. Demonstrating generality, we successfully designed probes for hydrogen peroxide (H2O2) and hydrogen sulfide (H2S). We envision a widespread adoption of the chemical platform for designing activatable NIR-II probes across diverse applications.