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Water-assisted laser desorption/ionization mass spectrometry for minimally invasive in vivo and real-time surface analysis using SpiderMass
摘要： Rapid, sensitive, precise and accurate analysis of samples in their native in vivo environment is critical to better decipher physiological and physiopathological mechanisms. SpiderMass is an ambient mass spectrometry (MS) system designed for mobile in vivo and real-time surface analyses of biological tissues. The system uses a fibered laser, which is tuned to excite the most intense vibrational band of water, resulting in a process termed water-assisted laser desorption/ionization (WALDI). The water molecules act as an endogenous matrix in a matrix-assisted laser desorption ionization (MALDI)-like scenario, leading to the desorption/ionization of biomolecules (lipids, metabolites and proteins). The ejected material is transferred to the mass spectrometer through an atmospheric interface and a transfer line that is several meters long. Here, we formulate a three-stage procedure that includes (i) a laser system setup coupled to a Waters Q-TOF or Thermo Fisher Q Exactive mass analyzer, (ii) analysis of specimens and (iii) data processing. We also describe the optimal setup for the analysis of cell cultures, fresh-frozen tissue sections and in vivo experiments on skin. With proper optimization, the system can be used for a variety of different targets and applications. The entire procedure takes 1–2 d for complex samples.
关键词： mass spectrometry,SpiderMass,real-time surface analysis,Water-assisted laser desorption/ionization,in vivo analysis
更新于2025-09-16 10:30:52
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Side Chain Optimization Remarkably Enhances the in Vivo Stability of <sup>18</sup> F-Labeled Glutamine for Tumor Imaging
摘要： Similar to glycolysis, glutaminolysis acts as a vital energy source in tumor cells, providing building blocks for the metabolic needs of tumor cells. To capture glutaminolysis in tumors, 18F-(2S,4R)4-Fluoroglutamine ([18F]FGln) and 18F-fluoroboronoglutamine ([18F]FBQ) have been successfully developed for Positron Emission Tomography (PET) imaging, but these two molecules are lack of stability, resulting in undesired yet significant bone uptake. In this study, we found that [18F]FBQ-C2 is a stable Gln PET tracer by adding two more methylene groups to the side chain of [18F]FBQ. [18F]FBQ-C2 was synthesized with a good radiochemical yield (RCY) of 35% and over 98% radiochemical purity. [18F]FBQ-C2 showed extreme stability in vitro, and no defluorination was observed after 2 hours in phosphate buffered saline at 37 °C. The competitive inhibition assay results indicated that [18F]FBQ-C2 enters cells via system ASC and N, similar to natural glutamine, and can be transported by tumor-overexpressed ASCT2. PET imaging and biodistribution results indicated that [18F]FBQ-C2 is stable in vivo with low bone uptake (0.81±0.20%ID/g) and can be cleared rapidly from most tissues. Dynamic scan and pharmacokinetic studies using BGC823-xenograft-bearing mice revealed that [18F]FBQ-C2 accumulates specifically in tumors, with a longer half-life (101.18±6.50 min) in tumor tissues than in other tissues (52.70±12.44 min in muscle). Biodistribution exhibits a high tumor-to-normal tissue ratio (4.8±1.7 for muscle, 2.5±1.0 for the stomach, 2.2±0.9 for the liver and 17.8±8.4 for the brain). In conclusion, [18F]FBQ-C2 can be used to perform high-contrast Gln imaging of tumors and can serve as a PET tracer for clinical research.
关键词： positron emission tomography,Glutamine,boramino acid,in vivo stability
更新于2025-09-12 10:27:22
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Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells Labeled with Mn2+ and Gd3+ Co-doped CuInS2-ZnS Nanocrystals for Multi-modality Imaging in Tumor Mice Model
摘要： Mesenchymal stem cells (MSCs) therapy has recently received profound interest as a targeting-platform in cancer-theranostics due to inherent tumor-homing abilities. However, the terminal tracking of MSCs-engraftment by fluorescent in situ hybridization, immuno-histochemistry and flow-cytometry techniques to translate into clinics are still challenging due to dearth of inherent MSC-specific markers and FDA-approval for genetic-modifications of MSCs. To address this challenge, a cost-effective non-invasive imaging technology based on multi-functional nanocrystals (NCs) with enhanced-detection sensitivity, spatial-temporal resolution, deep-tissue diagnosis is needed to be developed to track the transplanted stem cells. A hassle-free labeling of human umbilical-cord Wharton’s-Jelly (WJ)-derived MSCs with Mn2+ and Gd3+ co-doped CuInS2-ZnS (CIS-ZMGS) NCs has been demonstrated in 2 h without requiring electroporation process or transfection agents. It has been found that, WJ-MSCs labeling did not affect their multi-lineage differentiation (adipocyte, osteocyte, chondrocyte), immuno-phenotypes (CD44+, CD105+, CD90+), protein (β-actin, vimentin, CD73, α-SMCA) and gene-expressions. Interestingly, CIS-ZMGS-NCs labeled WJ-MSCs exhibit near-infrared fluorescence (NIR) with quantum yield (QY) of 84%, radiant intensity ~3.999 x 1011 (p/sec/cm2/sr)/(μW/cm2), magnetic relaxivity (longitudinal r1=2.26 mM-1s-1, transverse r2=16.47 mM-1s-1) and X-ray attenuation (78 HU) potential for early non-invasive multi-modality imaging of a subcutaneous-melanoma in B16F10-tumor-bearing C57BL/6-mice in 6 h. The ex vivo imaging and inductively-coupled plasma mass-spectroscopy (ICP-MS) analyses of excised organs along with confocal-microscopy and immuno-fluorescence of tumor results also significantly confirmed positive-tropism of CIS-ZMGS-NCs labeled WJ-MSCs in the tumor-environment. Hence, we propose the magneto-fluorescent CIS-ZMGS-NCs labeled WJ-MSCs as a next-generation nano-bioprobe of three commonly used imaging-modalities for stem cells-assisted anti-cancer therapy and tracking tissue/organ regenerations.
关键词： multi-modality imaging,stem cell labeling,CuInS2-ZnS,in vivo tracking,microwave refluxing
更新于2025-09-12 10:27:22
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In Vivo Reflection-Mode Photoacoustic Microscopy Enhanced by Plasmonic Sensing with an Acoustic Cavity
摘要： Relying on high-sensitivity refractive index sensing and a highly constrained evanescent field of surface plasmon resonance (SPR), broadband photoacoustic (PA) pressure transients were measured using an SPR sensor instead of routinely used piezoelectric ultrasonic transducers. An acoustic cavity made from stainless steel and having a designed ellipsoidal inner surface redirected laser-induced PA waves from the PA excitation spot to the SPR sensor. By incorporating the SPR sensor with the acoustic cavity, we developed optical-resolution photoacoustic microscopy (OR-PAM) with multiple advantages, including reflection-mode signal capture, improved PA detection sensitivity, increased PA spectral bandwidth as broad as ～98 MHz, and micrometer-scale lateral resolution. This allowed label-free volumetric PA imaging of vasculature in not only the thin ear but also the thick forelimb of living mice. With these combined advantages, our OR-PAM system potentially offers more opportunities for biomedical investigation, for example, when studying microcirculations in the eye and cortex.
关键词： reflection mode,high sensitivity,surface plasmon resonance,in vivo vascular imaging,photoacoustic imaging
更新于2025-09-12 10:27:22
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Application of quantum dot‐based biosensor to the conjunctiva in Wistar albino rats
摘要： This study was conducted to get an idea about the distribution of the lymphatic fluid in conjunctiva throughout the body parts. For this purpose, Qdot655 (QD), fluorescence nanoparticles, spread onto the conjunctiva were used on Wistar albino rats. Drainage of QD particles from conjunctiva was followed up via fluorescence images at different hours on body parts such as eye, ears, forearms, hind legs and tails. The first fluorescence signals within the 30th minutes following administration of QD were observed in the nasal region and the anterior extremities. Whereas within 60th minutes following QD spread, fluorescent signals were obtained from the ears, forearms, hind legs and tail of the female and male rats.
关键词： rat,conjunctiva,quantum dot,biosensor,in vivo imaging,eyelid lymphatics
更新于2025-09-11 14:15:04
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Noninvasive In Vivo Imaging in the Second Near-Infrared Window by Inorganic Nanoparticle-Based Fluorescent Probes
摘要： The fluorescence imaging in the second near-infrared window (NIR-II, 1000-1700 nm) has emerged as a new method for in vivo imaging and attracted considerable attention in the past decade. Owing to the suppressed photon scattering and diminished autofluorescence, in vivo fluorescence imaging in NIR-II window can afford deep tissue penetration depth with high clarity. Inorganic nanoparticle-based fluorescent probes in NIR-II window have greatly prospered the field into a development stage because of their superior traits, including adjustable emission covering the whole NIR-II window, and abundant surface functional groups that facilitate chemical modification and bioconjugation, etc. In this feature article, we introduce the unique imaging performance of the NIR-II optical window, and highlight the latest development of noninvasive biological fluorescent imaging in NIR-II window using inorganic nanoparticle-based probes. A perspective on the challenge and future direction of inorganic nanoparticle-based NIR-II probes is also discussed.
关键词： noninvasive imaging,NIR-II,inorganic nanoparticles,fluorescence imaging,in vivo imaging
更新于2025-09-11 14:15:04
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Quantitative and qualitative evaluation of recovery process of a 1064?nm laser on laser-induced skin injury: <i>in vivo</i> experimental research
摘要： Laser biological effects are a hot topic in laser medicine. In this study, to explore the quantitative biological effect of laser-induced wound healing and to provide guidance for expanding the clinical application of laser therapy, the injury effects and repair characteristics of skin tissue are studied through infrared laser irradiation of the skin of miniature pigs. Live pig skin was irradiated at multiple spots one time by using a grid-array method with a 1064 nm laser at different power outputs. The skin injury reaction was observed immediately after laser irradiation from low to high doses. The incidence of skin injury was calculated quantitatively. The healing and pathological changes after laser-induced skin injury were observed dynamically within 6 h and for 28 d after laser irradiation. With the increase of irradiation dose, laser-induced skin injuries ranging from mild to severe appeared in turn. The damage threshold of laser irradiation ED50 is 47.4 J cm?2 with the laser; from 3 d to 28 d after irradiation, the pathological results showed that wound healing tended to be different in all groups, but this trend weakened with the increase in laser irradiation intensity. With the increased irradiation dose, skin injury appears as different types of injury plaques, ranging from mild to severe. Skin injury is worsened and the tissue repair trend is weakened with the increase in laser irradiation dose, producing a good dose-effect and time-effect relationship.
关键词： laser-induced skin wound,infrared (IR) laser,wound healing,in vivo laser experiment
更新于2025-09-11 14:15:04
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In Vivo Two-photon Calcium Imaging in Dendrites of Rabies Virus-labeled V1 Corticothalamic Neurons
摘要： Monitoring neuronal activity in vivo is critical to understanding the physiological or pathological functions of the brain. Two-photon Ca2? imaging in vivo using a cranial window and speci?c neuronal labeling enables real-time, in situ, and long-term imaging of the living brain. Here, we constructed a recombinant rabies virus containing the Ca2? indicator GCaMP6s along with the ?uorescent protein DsRed2 as a baseline reference to ensure GCaMP6s signal reliability. This functional tracer was applied to retrogradely label speci?c V1–thalamus circuits and detect spontaneous Ca2? activity in the dendrites of V1 corticothalamic neurons by in vivo two-photon Ca2? imaging. Notably, we were able to record single-spine spontaneous Ca2? activity in speci?c circuits. Distinct spontaneous Ca2? dynamics in dendrites of V1 corticothalamic neurons were found for different V1–thalamus circuits. Our method can be applied to monitor Ca2? dynamics in speci?c input circuits in vivo, and contribute to functional studies of de?ned neural circuits and the dissection of functional circuit connections.
关键词： Corticothalamic projection,In vivo Ca2? imaging,Two-photon microscopy,Primary visual cortex,Neural circuit tracing,Cranial window,Rabies virus,Dendrite
更新于2025-09-11 14:15:04
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[IEEE 2018 IEEE International Conference on Robotics and Automation (ICRA) - Brisbane, QLD (2018.5.21-2018.5.25)] 2018 IEEE International Conference on Robotics and Automation (ICRA) - Design and Test of an In-Vivo Robotic Camera Integrated with Optimized Illumination System for Single-port Laparoscopic Surgery
摘要： This paper proposes a novel in-vivo robotic laparoscopic camera design with an optimized illumination system, which is a crucial component for achieving high imaging quality. The robotic camera design with three extendable wings can reserve sufficient on-board space to harbor the optimized illumination system without affecting the compactness of the camera. We contribute a freeform optical lens design method and develop three miniature optical lenses for the LEDs to achieve greater than 95% illumination uniformity, greater than 14, 000 lx illuminance on a target plane with a distance of 100 mm, and greater than 89% optical efficiency. The prototype is implemented and experimentally tested, which demonstrates great performance of the in-vivo robotic laparoscopic camera and the significance of the optimized illumination system.
关键词： single-port laparoscopic surgery,freeform optical lens design,in-vivo robotic laparoscopic camera,optimized illumination system
更新于2025-09-11 14:15:04
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Dynamic and non-contact 3D sample rotation for microscopy
摘要： Precise sample orientation is crucial for microscopy but is often performed with macroscopic tools and low accuracy. In vivo imaging of growing and developing samples even requires dynamic adaptation of the sample orientation to continuously achieve optimal imaging. Here, we present a method for freely positioning a sample in 3D by introducing magnetic beads and applying a magnetic field. We demonstrate magnetic orientation of fixed mouse embryos and artemia, and live zebrafish embryos and larvae on an epi-fluorescence microscope and on a light-sheet system for optimal imaging.
关键词： magnetic beads,microscopy,3D positioning,sample orientation,in vivo imaging
更新于2025-09-10 09:29:36