An ultralow-concentration, red-emissive dual-responsive fluorescent probe for real-time mitochondrial viscosity/polarity imaging.

Cellular imaging is a pivotal strategy for unraveling complex biological processes, disease mechanisms, and drug responses, wherein benzimidazole-acrylonitriles are emerging as promising yet underexplored fluorogenic scaffolds for advanced imaging and therapeutic applications. In the present study, five benzimidazole-acrylonitrile conjugates bearing nitrogen-rich heterocycles were synthesized and systematically investigated for their photophysical characteristics and aggregation-induced emission (AIE) behaviour. The synthetic strategy involves a stepwise condensation of o-phenylenediamine with ethyl cyanoacetate, followed by an L-proline-catalyzed condensation of the resulting intermediate with nitrogen-rich arylaldehydes. The molecular structures of the synthesized compounds were confirmed by 1H and 13C NMR spectroscopy and high-resolution mass spectrometry (HRMS); the structure of compound 5 was further unambiguously established by single-crystal X-ray diffraction analysis. Optical studies revealed distinct absorption and emission features attributable to the hybridized local and charge transfer excited state (HLCT), along with pronounced aggregation-induced emission (AIE) behavior in THF/water mixtures. The observed AIE characteristics were further supported by scanning electron microscopy (SEM) and dynamic light scattering (DLS), which revealed the formation of well-defined aggregated morphologies. All compounds demonstrated classic molecular-rotor-type fluorescence enhancement with increasing viscosity using a DMSO-glycerol mixture and showed distinct pH-responsive emission governed by protonation dynamics within the biological pH range, highlighting their potential as robust probes for visualizing intracellular heterogeneity and acidic microenvironments. Density functional theory (DFT) calculations provided complementary insights into the electronic structure and optical transitions. Remarkably, compound 1 exhibited efficient cytoplasmic localization in live HeLa cells, demonstrating its potential utility as a fluorescent bioimaging probe. Collectively, these findings establish benzimidazole-acrylonitrile conjugates as a new class of AIE-active luminogens with promising applications in precision bioimaging, tumor diagnostics, and theranostic platforms.

Yellow-emissive Nd-doped carbon quantum dots for ratiometric detection of anthrax biomarker with applications in food analysis and cellular imaging.

Cysteine (Cys), as one of the essential amino acids in humans, plays critical roles in numerous physiological processes, and its abnormal levels are closely associated with various diseases. Therefore, rapid and sensitive detection of Cys is of great significance for early disease diagnosis and mechanistic studies. In this work, we developed a novel near-infrared (NIR) fluorescent "turn-on" probe, NA-XL, featuring a large Stokes shift (177 nm) for highly efficient detection of the biothiol Cys. The probe incorporates a naphthalimide fluorophore and an acrylate group that serves as both the Cys recognition site and fluorescence quencher. The fluorescence quenching of NA-XL is predominantly governed by strong low-frequency vibrations. Upon selective reaction with Cys, this vibration-mediated quenching process is effectively suppressed, leading to a pronounced NIR fluorescence emission at 729 nm. The probe demonstrates excellent selectivity and sensitivity, achieving a low detection limit (LOD) of 10.7 nM. Furthermore, NA-XL exhibits outstanding responsiveness to both exogenous and endogenous Cys in cellular imaging, enabling real-time monitoring of Cys dynamics. These findings highlight its potential for biomedical research applications.

High-performance liquid chromatography enables purification of red-emitting carbon dots for cellular imaging.

Photoluminescence organic wide-band gap semiconductors investigated for optoelectronic applications and cellular imaging

A peptide-functionalized quantum Dots/MOF nanosheets fluorescence biosensor for glutathione sensing and cellular imaging.

Ongoing Clinical Challenges in Nerve Surgery (Nerve SPACE 2025).

Alkoxy chain engineering in diphenylamine-based fluorophores: Achieving red-shifted near-infrared aggregation-induced emission for cellular imaging.

Lipid droplet-targeted fluorescent probes for polarity sensing in Ferroptosis cellular models and in vivo zebrafish.

Donor-acceptor conjugation engineering boosts polarity sensitivity: dual-channel monitoring of lipid metabolic flux.

Imaging flow cytometry using Fourier light-field microscopy enables high-throughput three-dimensional cellular imaging, capable of capturing thousands of events per second. However, volumetric reconstruction speed remains orders of magnitude slower than the acquisition speed. The current state of art uses Richardson-Lucy algorithm, restricted to just 5-10 reconstructed events per second with GPU acceleration. This limitation hinders real-time applications such as cell sorting and thus has bottlenecked the widespread adoption of 3D imaging flow cytometry. We introduce patch deconvolution, the first training-free algorithm compatible with the Richardson-Lucy framework that significantly accelerates convergence, achieving over 100-200 reconstructions per second on standard GPUs, a 20- to 40-fold improvement over Richardson-Lucy. Validated on both simulated and experimental data sets, patch deconvolution achieves reconstruction quality comparable to Richardson-Lucy in both static and flow data. This supports rapid cell sorting based on spatial features and enables advanced applications, such as detecting rare spatial events in large cell populations, which would otherwise be indistinguishable in traditional flow cytometry.

Label-free Raman microspectroscopic cellular imaging and untargeted UPLC-HRMS metabolomics of apple branch canker progression based on chemometrics and machine learning

A degradable NIR Ratiometric probe for HClO monitoring from cellular imaging to smart platforms.

The development of anisotropic gold nanostructures supporting localized surface plasmon (LSP) resonances in the near-infrared (NIR) biological window is of great interest for diagnostic and therapeutic nanotechnologies. Here, we report gold open-shell nanoparticles (AuOSNs), a symmetry-broken nanoshell architecture exhibiting strong NIR surface-enhanced Raman scattering (SERS) activity. AuOSNs were fabricated via a surfactant-free strategy combining bottom-up silica sphere assembly with a simple top-down gold deposition process, without using highly cytotoxic surfactants such as cetyltrimethylammonium bromide (CTAB). Boundary element method (BEM) simulations revealed that the asymmetric open-shell geometry induces NIR LSP resonances with pronounced electromagnetic field localization near the opening edges, depending on excitation configuration. Consistent with these predictions, extinction spectra of AuOSNs dispersed in water showed an LSP resonance peak at ~793 nm, close to the 785 nm excitation wavelength for SERS. In aqueous dispersion, AuOSNs modified with 4-mercaptobenzoic acid (4-MBA) exhibited strong SERS activity with enhancement factors of ~106. Furthermore, polyethylene glycol (PEG)-modified MBA/AuOSNs showed negligible cytotoxicity in vitro. SERS imaging confirmed that PEG/MBA/AuOSNs enable visualization of HeLa cells via characteristic MBA SERS signals. These results demonstrate that surfactant-free AuOSNs provide a biocompatible platform for NIR-excited SERS sensing and cellular imaging, highlighting their potential in plasmonic bioimaging applications.

The enhancement of efficacy via the optimization of cancer drug ratios in lipid–polymer hybrid nanocapsules (LPHNs), including doxorubicin (DOX) and paclitaxel (PTX), at diverse ratios, represents a viable approach for optimizing cancer therapy results. This study examined four specific (DOX: PTX) ratios, 20:80 (C1), 40:60 (C2), 60:40 (C3), and 80:20 (C4), to determine the best formulation and develop a dual-loaded fluorescent DOX-PTX nanocapsule with controlled release features for targeting breast cancer cells. This nanocapsule (NC), functionalized with folic acid (FA) and fluorescein isothiocyanate (FITC), exhibited accurate targeting abilities and in vitro visibility, indicating its use in individualized cancer treatment. The structural and physicochemical properties were evaluated via DLS, FTIR, XRD, PL spectroscopy, FESEM, and TEM. The cytotoxicity assay determined the average IC50 values for the MCF-7 cell line at 24 and 48 h, along with the cytotoxicity data presented in four sets, which were compared with those of the free drug against the MCF-7 cancer cell line. The encapsulation and release properties confirmed consistent drug loading and extended drug delivery. Moreover, the advancement of controlled release holds significant promise for enhancing its effectiveness. Single-cell gel electrophoresis (SCGE) demonstrated the pronounced genotoxic effects of LPHNc, which was corroborated by cellular imaging, indicating effective absorption and distribution. The optimized drug concentration induced prominent DNA damage, G2/M phase arrest, and a notable sub-G1 population, confirming apoptosis via cell cycle analysis. These findings highlight the efficacy of LPHNc in inducing genotoxicity, disrupting proliferation, and causing cell death with a steady slope.

Abstract Background Nipple-sparing mastectomy (NSM) followed by breast reconstruction is now widely recognized as a safe technique, including in patients with a history of radiotherapy or chemotherapy, regardless of tumor size or axillary status, except in cases of inflammatory breast cancer or direct involvement of the nipple-areola complex on imaging or clinical examination. The main oncological concern is the potential persistence of tumor remnants in the residual subareolar breast tissue. At least 10% of patients undergo a second surgery to excise this tissue. Such reinterventions may increase patient anxiety, negatively impact body image, raise healthcare costs, and delay adjuvant treatments. A new generation of ultra-fast confocal microscopes (UFCM) enables high-resolution imaging of fresh tissue in a shorter time than frozen section analysis, potentially guiding intraoperative assessment and avoiding reoperation [1-4]. Methods The CAMELIA project is a prospective, non-interventional, ex vivo clinical study analyzing retroareolar margins after NSM, imaged using UFCM and interpreted by pathologists and surgeons. The examined tissue is preserved for subsequent conventional histology (HES). At this stage, UFCM results do not influence surgical management. The primary objective is to use UFCM to provide an accurate intraoperative diagnosis of retroareolar tissue. Accuracy, sensitivity, and specificity are calculated based on the proportion of images correctly interpreted compared to definitive histology on corresponding HES slides, the reference standard. Results Since October 2024, 40 out of 194 patients who underwent Nipple-Sparing Mastectomy (NSM) have been included in this study. Of these, 8 had prophylactic mastectomies and 32 had therapeutic interventions. There were no bilateral procedures, and 16 patients underwent surgery after neoadjuvant chemotherapy. Importantly, four of these patients had positive margins. Three pathologists participated in the image analysis: two seniors (PT1 and PT2) and one junior (PT3). They analyzed 20 out of 40 images. Both PT1 and PT3 have experience with Ultra-Focalized Cancer Mammography (UFCM). Additionally, an experienced UFCM surgeon interpreted all 40 UFCM images. Images were categorized into four groups: deferred diagnosis, cancer, no cancer, or suspicious. For the first 20 cases, the pathologists' concordance rate ranged from 59% to 75%. Specifically: PT1 classified 2 out of 3 positive margins as suspicious, PT2 deferred diagnosis for 3 cases and classified 1 out of 3 as suspicious. PT3 classified 1 out of 3 positive margins as suspicious. The surgeon's concordance rate for the first 40 cases was 72.5%, with one positive margin identified out of four existing positive margins. On average, the intraoperative image acquisition time was 16 minutes. Readers rated the quality of these images at 3.4/5. The average image interpretation time across all specialists was 1 minute and 30 seconds. Conclusions Expected outcomes include high accuracy, sensitivity, and specificity. These results will provide proof of concept for cellular imaging of the retroareolar margin during NSM, with the aim of completing tumor resection if necessary and avoiding reoperation.

Functionalized dibenzothiophene‐S,S‐dioxides (DBTO) have emerged as a versatile class of heteroaromatic scaffolds with remarkable optoelectronic and photophysical properties. Their rigid π ‐conjugated framework, combined with the strong electron‐withdrawing nature of the sulfone group, facilitates precise tuning of frontier molecular orbitals, enhancing charge transport and luminescence. This review presents a comprehensive exploration of synthetic strategies aimed at diversifying the structural and electronic landscape of functionalized DBT‐SO 2 derivatives. Key methodologies include regioselective bromination, directed lithiation, Suzuki–Miyaura and Stille cross‐coupling reactions, as well as oxidative and reductive modifications to introduce electron‐donating (or) electron‐withdrawing substituents. By fine‐tuning the substitution pattern and conjugation length, a diverse set of DBTO based molecules was synthesized with tailored optical bandgaps and charge transport properties. The resulting compounds exhibit tunable absorption and emission properties, high photostability, and strong fluorescence quantum yields, making them promising candidates for multifunctional applications. Detailed spectroscopic, electrochemical, and thermal characterizations reveal structure–property relationships critical for optimizing performance in organic light‐emitting diodes (OLEDs), organic field‐effect transistors (OFETs), and fluorescence‐based bioimaging platforms. Furthermore, select derivatives demonstrate excellent photostability and biocompatibility, enabling their use as fluorescent probes for cellular imaging. Their strong absorption in the UV–visible region, combined with deep‐blue to green emission and low cytotoxicity, underscores their promise in biomedical applications. Overall, this work provides a systematic framework for the molecular design and functional optimization of DBT‐SO 2 ‐based materials. The results emphasize how structural engineering can be leveraged to unlock multifunctional performance, bridging the fields of organic electronics and bioimaging. These findings open new avenues for the development of high performance, tunable organic materials based on the DBTO scaffold.

Sensory deprivation leads to extensive cortical plasticity, but the impact of enhanced sensory experience on the mature cortex remains poorly understood. Here, we examine how visually evoked activity in cortical circuits is shaped by repeated exposure to varied stimuli. The most prominent pattern of visually evoked activity in mouse primary visual cortex, beta oscillations (15-30Hz), arises from brief events of neural synchrony with a characteristic pattern of laminar propagation and relies on the activity of somatostatin-expressing (SST) GABAergic interneurons. We find that visually evoked beta activity is initially weak but robustly potentiated by repeated exposure to a diverse visual stimulus set, leading to enhanced recruitment of cortical neurons by these rhythmic network events. Cellular imaging further reveals that visual experience leads to increased visual responses in SST interneurons and suppressed responses in vasoactive intestinal peptide-expressing (VIP) GABAergic interneurons. In association with this rebalancing of inhibitory circuits, visual experience enhances visual selectivity in nearby pyramidal neurons. Visual experience thus selectively reorganizes adult dendrite-targeting inhibitory circuits, promoting network synchrony and enhancing sensory encoding by cortical excitatory projection neurons.

Understanding single-cell neuronal activity across multiple brain regions in the context of ethologically relevant behaviors is a major goal in systems neuroscience. We have engineered the Cortex Camera Array Microscope (CortexCAM), integrating four miniaturized fluorescence imaging microscopes to simultaneously capture cellular activity from contiguous fields of view spanning over 48 mm 2 of the dorsal cortex. The CortexCAM is capable of imaging > 9000 individual neurons across much of the primary and secondary motor, somatosensory, visual, retrosplenial, and association cortices across both hemispheres of the dorsal cortex. The compact nature of the CortexCAM allows integration into a passive mechanical gantry system to form the mobile CortexCAM. The mobile CortexCAM allows volitional control of translational motion (x, y) and rotational motion (yaw) in physical behavior arenas. We then use the mobile CortexCAM to perform cortex-wide cellular resolution imaging in freely locomoting mice performing alternating choice tasks, as well as during social interactions. Thus the CortexCAM allows studying cortex-wide cellular dynamics in behaviors that cannot be achieved in headfixed settings.