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- 实验方案
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High-resolution cost-effective compact portable inverted light microscope
摘要： Commercial high-resolution optical microscopes are essential for microscopy imaging; however, they are expensive and bulky, which limits their use in point-of-care devices, resource-limited areas, and real-time imaging of a sample in a large apparatus. In this study, we report a novel compact (10 cm × 5 cm × 5 cm, without the light source) lightweight (≤0.5 kg) submicron-resolution inverted optical microscope at low cost (≤$ 300). Our technique utilises the proximity of the image sensor to a commercial microscope objective lens for compactness of the microscope. The use of an image sensor with a small pixel size helps to reduce the information loss, which provides high-resolution images. Moreover, our technique offers a freedom to tailor the design of microscope according to the required resolution, cost, and portability for specific applications, which makes it a suitable candidate for affordable point-of-care devices. Images of several micron-to-submicron scale patterns and spherical beads are acquired to observe the resolution and quality of the images obtained using our microscope. In addition, we demonstrate the applications of our microscope in various fields such as recording of high-speed water micro-droplet formation inside a microfluidic device, high-resolution live cell imaging inside an incubator, and real-time imaging of crack propagation in a sample under stretching by a material testing system (MTS). Therefore, this portable and inexpensive microscope provides the essential functionalities of a bulky expensive high-performance microscope at a lower cost.
关键词： portable inverted microscope,microfluidics,discrete wavelet transform,submicron-scale imaging,industrial microscope,Cell imaging
更新于2025-09-04 15:30:14
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A machine learning approach for online automated optimization of super-resolution optical microscopy
摘要： Traditional approaches for finding well-performing parameterizations of complex imaging systems, such as super-resolution microscopes rely on an extensive exploration phase over the illumination and acquisition settings, prior to the imaging task. This strategy suffers from several issues: it requires a large amount of parameter configurations to be evaluated, it leads to discrepancies between well-performing parameters in the exploration phase and imaging task, and it results in a waste of time and resources given that optimization and final imaging tasks are conducted separately. Here we show that a fully automated, machine learning-based system can conduct imaging parameter optimization toward a trade-off between several objectives, simultaneously to the imaging task. Its potential is highlighted on various imaging tasks, such as live-cell and multicolor imaging and multimodal optimization. This online optimization routine can be integrated to various imaging systems to increase accessibility, optimize performance and improve overall imaging quality.
关键词： machine learning,multicolor imaging,online automated optimization,live-cell imaging,super-resolution optical microscopy,multimodal optimization
更新于2025-09-04 15:30:14
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A novel OFF-ON-OFF fluorescence probe based on coumarin for Al3+ and F? detection and bioimaging in living cells
摘要： A novel fluorescence probe L2 based on coumarin has been designed and synthesized. The probe L2 can be used for relay recognition of metal ions Al3+ and anion F? in the aqueous HEPES buffer (0.05 M, pH=7.4), and build a OFF-ON-OFF detection system. The probe showed high selectivity and sensitivity to target ions in the process of relay recognition, and the corresponding detection limit could be as low as 0.014 μM (Al3+) and 0.03 μM (F?). Besides, the geometry optimizations of probe L2 and [L2+Al3+] complex were carried out using the Gaussian 16 program based on DFT, and the identification mechanism of the probe was also discussed by the mass spectrometry and theoretical calculations. Moreover, the probe have also been successfully applied to detection of target ions in living cells.
关键词： Al3+,coumarin,Fluorescence probe,cell imaging,F?
更新于2025-09-04 15:30:14
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Deep-red fluorogenic probe for rapid detection of nitric oxide in Parkinson’s disease models
摘要： Nitric oxide (NO), as one of important gaseous signaling molecule in human body, has been associated with a variety of physiologic processes. However, excessive production of NO has been profoundly implicated in the pathogenesis of neurodegenerative disorders in particular, Parkinson's disease (PD). Therefore, accurate and facile detection of NO is of great significance for investigating its functions in PD, and the subsequent diagnosis and/or treatment. Herein, we developed a deep-red fluorogenic probe (BT-NH), with high sensitivity and good selectivity to detect NO, which was successfully used to visualize exogenous/endogenous NO level in living cells, and further applied to in vitro and in vivo PD models.
关键词： Parkinson’s disease,Drosophila,live cell imaging,nitric oxide,fluorogenic probe
更新于2025-09-04 15:30:14
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Gadolinium Doping Enhances the Photoacoustic Signal of Synthetic Melanin Nanoparticles: A Dual Modality Contrast Agent for Stem Cell Imaging.
摘要： In this paper we show that gadolinium-loaded synthetic melanin nanoparticles (Gd(III)-SMNPs) exhibit up to a 40-fold enhanced photoacoustic signal intensity relative to synthetic melanin alone and higher than other metal-chelated SMNPs. This property makes these materials useful as dual labelling agents because Gd(III)-SMNPs also behave as magnetic resonance imaging (MRI) contrast agents. As a proof-of-concept, we used these nanoparticles to label human mesenchymal stem cells (hMSCs). Cellular uptake was confirmed with bright field optical and transmission electron microscopy. The Gd(III)-SMNP labeled stem cells continued to express the stem cell surface markers CD73, CD90, and CD105 and proliferate. The labeled stem cells were subsequently injected intramyocardially in mice, and the tissue was observed by photoacoustic and MR imaging. We found the photoacoustic signal increased as cell number increased (R2 = 0.96) indicating that such an approach could be employed to discriminate between stem cells populations with a limit of detection of 2.3 x 104 cells in in vitro tests. This multimodal photoacoustic/MRI approach combines the excellent temporal resolution of photoacoustics with the anatomic resolution of MRI.
关键词： Photoacoustic Signal,Stem Cell Imaging,Gadolinium,Dual Modality Contrast Agent,Synthetic Melanin Nanoparticles
更新于2025-09-04 15:30:14
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A ratiometric fluorescent probe for hydrazine detection with large fluorescence change ratio and its application for fluorescence imaging in living cells
摘要： Hydrazine and its derivatives are one of the most important raw chemical materials that have extensive utilization in many areas. It is also widely applied as pharmaceutical derivatives, such as the antipsychotic drug, anti-infective drug, and antitumor drug. However, hydrazine is a highly toxic reagent which can bring severe damage to human and animals. Herein, we have reported a new benzothiazole substituted naphthalene based probe for hydrazine with huge fluorescence change ratio. The fluorescence change ratio at F390/F509 is up to 200-fold in the presence of hydrazine in 60 min. The probe exhibited negligible cytotoxicity toward HeLa cells and was applied for hydrazine detection successfully in HeLa cells.
关键词： Fluorescent probe,ratiometric,cell imaging,hydrazine detection
更新于2025-09-04 15:30:14
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Fluorogenic “Photoclick” Labeling and Imaging of DNA with Coumarin-fused Tetrazole in Vivo
摘要： Photoclickable fluorogenic probes will enable visualization of specific biomolecules with precise spatiotemporal control in their native environment. However, the fluorogenic tagging of DNA with current photocontrolled clickable probes is still challenging. Herein, we demonstrated the fast (19.5 ± 2.5 M-1 s-1) fluorogenic labeling and imaging of DNA in vitro and in vivo with rationally designed coumarin-fused tetrazoles under UV LED photoirradiation. With a water-soluble, nuclear-specific coumarin-fused tetrazole (CTz-SO3), the metabolically synthesized DNA in cultured cells was effectively labeled and visualized, without fixation, via “photoclick” reaction. Moreover, the photoaclickable CTz-SO3 enabled the real-time, spatially controlled imaging of DNA in live zebrafish.
关键词： DNA,cell imaging,fluorogenic,metabolic labeling,photoclick
更新于2025-09-04 15:30:14