Jove-Journal of Visualized Experiments最新文献

{"title":"3D Cine Magnetic Resonance Imaging of Respiratory Motion in Mechanically Ventilated Mice and Rats.","authors":"Myrte Wennen, Wout Claassen, Nick van Huis, Ruslan Garipov, Lindy Alles, Leo Heunks, Coen Ottenheijm, Bram Coolen, Gustav Strijkers","doi":"10.3791/67474","DOIUrl":"https://doi.org/10.3791/67474","url":null,"abstract":"<p><p>The pathophysiology of diaphragm dysfunction in mechanically ventilated patients has yet to be fully understood, and adequate animal models are required to accommodate further research. Mechanical ventilation settings, such as the level of positive end-expiratory pressure (PEEP), play a crucial role. The goal was to develop a method to image 3D thoracic movement during mechanical ventilation of mice and rats at different respiratory pressure settings. Rats (Wistar) and mice (C57BL/6) were anesthetized with a mix of ketamine, atropine, and dexmedetomidine. Anesthesia was maintained by continuous infusion through a peritoneal catheter. Next, a tracheostomy was performed to enable mechanical ventilation of the animals. Animals were placed in a 7T MR system while ventilated with an MR-compatible ventilator. 3D cine imaging of the thorax was conducted using a 3D gradient echo sequence with pseudo-spiral k-space filling. A navigator signal, generated by the slice selection rewinder gradient, was recorded every TR. Retrospective binning and reconstruction of the data in 12 respiratory cine time frames was performed using in-house developed software. We successfully visualized thoracic movement in 3D in both species during mechanical ventilation, enabling the investigation of changes in thorax geometry throughout the respiratory cycle at varying PEEP levels. We found that retrospective binning of respiratory frames was highly facilitated by the fixed respiration rate. The protocol presented here can be used to study cardiac and thoracic geometry and movement in mechanically ventilated mice and rats.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145239828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Multifunctional, Embedded-based, Bluetooth-enabled, Programmable, Biphasic-waveform Stimulator with Real-time Neural Signal Acquisition.","authors":"Paul V Milner, Philips Zachariah, Jijo Francis, Siji George, Adarsh V Parekkattil, Suni Jose, S Athithya, Caren Babu, Binoy Johnson, Loitongbam Surajkumar Singh, Shuma Adhikari","doi":"10.3791/68837","DOIUrl":"https://doi.org/10.3791/68837","url":null,"abstract":"<p><p>The study introduces a lightweight, Bluetooth-enabled deep brain stimulation (DBS) device that is designed specifically for preclinical research in small, freely moving animals. To address the current limitations in waveform flexibility and wireless control, we developed a compact, multifunctional stimulator capable of delivering programmable, charge-balanced biphasic waveforms while simultaneously acquiring neural signals. The system integrates off-the-shelf components for current regulation, signal amplification, and analog-to-digital conversion, all managed by a low-power microcontroller. The assembly includes key steps such as circuit integration on a double-sided 30 x 30 mm PCB, waveform programming via nRF Connect, and validation through saline and load resistance testing. In vitro assessments demonstrated reliable current output across varying load impedances, effective saline operation without waveform degradation, and high-fidelity neural signal recording with a signal-to-noise ratio exceeding 35 dB. These results confirm the device's suitability for closed-loop neuromodulation experiments and lay the groundwork for future translational studies in DBS therapy.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145239975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enabling High-Throughput Perovskite FET Research by a Customizable Automated FET Measurement Station.","authors":"Claas Wieland, Giovanni Ligorio, Emil List-Kratochvil","doi":"10.3791/68573","DOIUrl":"https://doi.org/10.3791/68573","url":null,"abstract":"<p><p>Perovskite-based thin-film field effect transistors (PeFETs) have yet to achieve the full theoretical potential of this promising class of materials. To bridge this gap, it is essential to develop and optimize novel perovskite compositions and fabrication techniques. Given the large variety of potential compounds as well as the manifold of influencing variables such as concentration, temperature, and choice of solvent, a high-throughput research approach is critical for efficient exploration and advancement. We present a flexible and customizable process that spans from substrate fabrication to device characterization. This process is enabled by and integrates photolithography for custom patterning, an automated measurement station for FET characterization, and automated data analysis. The automated measurement station is based on a multiplexer, which is connected to five measurement boards. The measurement boards are configured to measure one substrate with four devices each. Allowing the automated measurement of 20 devices with up to 5 different perovskite formulations. Using standardized testing procedures, the raw data is automatically analyzed to get the transfer and output characteristics of the PeFETs as well as key performance parameters like the threshold voltage, subthreshold swing, and the field effect mobility. The result is a systematically organized data pool with easily comparable data for different perovskite compositions or modifications in fabrication conditions.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Standardized Approach to Superior Mesenteric Artery Resection for Locally Advanced Pancreatic Cancer.","authors":"Carl-Stephan Leonhardt, Ulla Klaiber, Klaus Sahora, Oliver Strobel","doi":"10.3791/68465","DOIUrl":"https://doi.org/10.3791/68465","url":null,"abstract":"<p><p>Novel multimodal treatment strategies might enable curative-intent resection of locally advanced pancreatic cancer (LAPC). While venous resections are routinely performed, arterial resections remain part of individualized treatment strategies in a few expert centers. Historically, SMA involvement, by definition, precluded resectability. However, in selected patients with excellent response to preoperative treatment, arterial resection has been associated with satisfactory outcomes in high-volume centers with significant experience in extended and vascular resections. Even though arterial divestment is increasingly performed if technically possible, arterial resection is still required when the arterial wall cannot be preserved to achieve macroscopically complete resection, especially if the SMA is encased along with its branches in SMA segment 2. Usually, the SMV is also encased in these tumors, and combined arterial and venous resections are necessary. Here, we describe our approach to SMA resection and reconstruction after neoadjuvant treatment in a 64-year-old female patient with locally advanced pancreatic ductal adenocarcinoma (PDAC) and complete encasement of the SMA in segment 2. Critical steps in this situation are assessing the feasibility of SMA and SMV resection using an infracolic artery first approach, performing a wide Kocher maneuver to access the SMA from the right, and the Cattell-Braasch maneuver to enable tension-free direct end-to-end anastomoses.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Laser Micromachining for Polymer Surface Topography Design.","authors":"Izidor Straus, Gaia Kravanja, Luka Hribar, Raphael Kriegl, Mikhail Shamonin, Irena Drevenšek-Olenik, Matija Jezeršek, Gašper Kokot","doi":"10.3791/68126","DOIUrl":"https://doi.org/10.3791/68126","url":null,"abstract":"<p><p>Soft magnetoactive elastomers (MAEs) are smart materials that respond to external magnetic fields by dynamically altering their mechanical properties. They are composed of magnetically responsive microparticles embedded within a soft polymer matrix, exhibiting an effective shear modulus of up to 100 kPa. In recent decades, MAEs' bulk properties have been successfully exploited for applications such as dynamic vibration damping, vibration sensing, and actuation in soft robotics. Recent research has shifted to their surface properties, revealing promising results on tunable surface features such as roughness, adhesion, and wetting. Even the transport of small solid and fluid objects was demonstrated. The associated surface effects can be significantly enhanced through the precise engineering of surface topography. In this article, an efficient laser micromachining technique, with a resolution of 15 µm, is presented, which enables rapid prototyping of MAE surfaces. It allows the creation of various complex shapes and offers functionality beyond the one achievable with traditional molding techniques. Additionally, the approach is versatile and can be applied to any polymer that sufficiently absorbs the laser light. As an example, a lamellar surface micro-pattern fabrication process and its characterization by optical and scanning electron microscopies are shown. Its response to a magnetic field is demonstrated. The technique provides a flexible and fast solution for optimizing polymer surface design across a wide range of applications.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Real-Time Indocyanine Green Fluorescence Navigation in Difficult Laparoscopic Cholecystectomy.","authors":"Can-Hua Zhu, Yan-Na Chen, Jun-Hua Cen","doi":"10.3791/69001","DOIUrl":"https://doi.org/10.3791/69001","url":null,"abstract":"<p><p>Laparoscopic cholecystectomy (LC), with its advantages of minimal invasiveness and rapid recovery, has become the standard surgical approach for benign gallbladder diseases. However, even experienced surgeons cannot completely avoid bile duct injury (BDI), and the incidence of BDI during LC is 2-3 times higher than that of open surgery, making it the most common cause of iatrogenic BDI. Approximately 20% of BDIs require multiple surgeries, and about 0.8% eventually result in liver transplantation, significantly impacting patient safety and quality of life, while posing a major risk for medical disputes. Achieving real-time intraoperative visualization is crucial to preventing BDI, particularly in difficult cholecystectomy cases under inflammatory conditions. Real-time indocyanine green (ICG) fluorescence guidance during LC can enhance extrahepatic bile duct visualization and minimize the risk of bile duct injury. In this surgical protocol, an 83-year-old female patient, who had undergone percutaneous transhepatic gallbladder drainage (PTGBD) for acute suppurative cholecystitis 6 weeks prior, was admitted for LC. During the operation, 2.5 mg of ICG was intravenously administered 10 min before the skin incision. Twenty minutes after injection, the liver and common bile duct were clearly visualized under fluorescence imaging. As the dissection of Calot's triangle progressed, the gallbladder and cystic duct remained unstained due to stone impaction, creating a stark visual contrast. Under dynamic ICG guidance, Calot's triangle was meticulously dissected, and the cystic artery and cystic duct were sequentially ligated and divided. The gallbladder was successfully removed. The surgical field showed no bleeding, and fluorescence imaging confirmed no bile leakage, achieving visualized LC under severe inflammatory adhesions. The patient was discharged on the third postoperative day. Real-time ICG fluorescence navigation for LC is safe, effective, and minimally invasive, particularly suitable for difficult cases.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid Detection of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus by Multiplex Real-Time Fluorescent PCR.","authors":"Shu-Ni Li, Cheng-Yuan He, Yi-Na Zhao, Xin-Qiang Zhang, Ge Huang, Jin-Xin Lai, Zheng-Kang Li, Guang-Hua Li","doi":"10.3791/69004","DOIUrl":"https://doi.org/10.3791/69004","url":null,"abstract":"<p><p>Staphylococcus aureus (S.aureus) is a common Gram-positive human pathogen that causes community-acquired and nosocomial infections, with diverse clinical manifestations ranging from local superficial lesions and food poisoning to fatal systemic infections. The discovery of antibiotics significantly reduced the mortality rate, but the problem of drug resistance has since become increasingly prominent. Since the first identification of MRSA in 1960, this strain has emerged as a global public health threat. MRSA is a major pathogen of nosocomial infections, capable of causing various severe diseases such as endocarditis, chronic osteomyelitis, pneumonia, pyogenic arthritis, and bacteremia. Therefore, rapid and accurate detection of S. aureus and its drug resistance is crucial for guiding clinical treatment. Current routine detection methods for S.aureus and MRSA have significant limitations. The traditional bacterial culture method, serving as the \"gold standard\" for decades, can provide definitive species identification and drug sensitivity results, but the process is time-consuming, taking 48 to 72 h. Furthermore, this method is susceptible to contamination and relies on specialized laboratory facilities and skilled technicians. Serological testing achieves non-invasive diagnosis by detecting S.aureus antibodies in patient serum, but it cannot distinguish between active infections and past infections, nor can it identify drug-resistant strains (such as MRSA). This study focuses on developing a novel multiplex real-time fluorescence PCR detection method to overcome the above-mentioned limitations. This method designs specific primers and TaqMan fluorescent probes targeting the species-specific nuc gene of S.aureus and the mecA gene mediating methicillin resistance, enabling simultaneous amplification and detection of S.aureus and MRSA in a single reaction system. This technology greatly reduces the detection time, providing a rapid, accurate, and cost-effective solution for S.aureus and MRSA detection. This innovative approach greatly improves clinical diagnostic efficiency and facilitates the early implementation of targeted antibiotic therapy, making important contributions to controlling drug-resistant bacterial infections.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling Dysplastic and Functional Lung Alveolar Repair after Influenza Infection.","authors":"Sachin Gaurav, Arnav Sharma, Francisco Lobo, Terren K Niethamer","doi":"10.3791/69062","DOIUrl":"https://doi.org/10.3791/69062","url":null,"abstract":"<p><p>Viral infection causes both acute and long-term damage to the lung alveolus, the specialized tissue structure responsible for gas exchange between the cardiovascular system and the external environment. Influenza A virus (IAV) infection in mice represents a translational model for the response of the human lung to viral infection and induces both transient and persistent cell state changes in the alveolus. In some cases, aberrant cell states induced by viral injury that are not resolved over time may permanently impair the critical gas exchange function of the lung. This article demonstrates methods for intranasal infection of mice with A/PR/8/34 IAV and characterization of transient and long-lasting alterations to the cellular composition and tissue structure of damaged lungs. This model enables a detailed investigation into the molecular and cellular mechanisms underlying lung repair and chronic dysfunction. This approach also offers a platform for evaluating therapeutic interventions aimed at promoting effective lung regeneration and restoring respiratory function after viral injury.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Modified Co-Culture System for Understanding Granulosa-Theca Cell Interactions in the Bovine Ovary.","authors":"Anja Baufeld, Jens Vanselow","doi":"10.3791/68589","DOIUrl":"https://doi.org/10.3791/68589","url":null,"abstract":"<p><p>A reliable co-culture of granulosa (GCs) and theca cells (TCs) has the potential to facilitate a more comprehensive understanding of the interactions, signaling pathways, and substrate exchange between both cell compartments in the bovine ovarian follicle. Utilizing commercially available cell culture inserts, a reproducible co-culture model of bovine GCs and TCs was established. It was observed that primary theca and granulosa cells require considerably more time to attach to the insert membrane compared to cell lines. Initially, the TCs were seeded onto the inverted insert membrane, with a truncated tube serving as an inoculation container to prevent medium leakage. Following a 72 h incubation period, the insert was inverted, allowing the GCs to be cultured on the opposite side of the membrane. This co-culture was then subjected to an additional 6 days of incubation at 37 °C and 5% CO2, with media exchange every other day. The expression of marker genes indicated cell-type-specific patterns, with CYP17A1 being highly abundant in TCs and not expressed in GCs. Conversely, CYP19A1 showed high levels in GCs and only low levels in TCs. The hormone analysis supported the presence of a physiological co-culture system, as evidenced by the synthesis of estradiol. This modified co-culture model supports reproducible studies of paracrine signaling and substrate transport between the somatic cells of the follicle.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145239856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anticancer Effects of Key Constituents of Pileostegia tomentella via ROS-Mediated Classical Apoptosis in Non-Small Cell Lung Cancer Cells.","authors":"Wen Zhong, Leimin Jiang, Bing Qing, Xiumei Ma, Qingmei Deng, Guangyun Zhou, Jiangcun Wei","doi":"10.3791/69008","DOIUrl":"https://doi.org/10.3791/69008","url":null,"abstract":"<p><p>This study explores the anticancer potential of Pileostegia tomentella, with a particular focus on its ability to induce classical apoptosis in H1299 non-small cell lung cancer (NSCLC) cells through reactive oxygen species (ROS)-mediated pathways. Using a bioactivity-guided isolation approach, two active compounds, umbelliferone (UMB) and epi-Vogeloside, were identified as the major cytotoxic constituents. Both compounds exhibited significant inhibitory effects on cell viability and effectively triggered apoptotic cell death in H1299 cells. Mechanistic investigations demonstrated that treatment with UMB and epi-Vogeloside led to a marked increase in intracellular ROS levels, implicating oxidative stress as a key mediator. Western blot analysis revealed increased levels of cleaved caspase-3 and decreased expression of the anti-apoptotic protein Bcl-2, indicating activation of the intrinsic mitochondrial apoptotic pathway. Additionally, transcriptomic profiling showed widespread alterations in gene expression, further supporting disruption of cellular homeostasis. These results suggest that Pileostegia tomentella and its active constituents hold promise as potential therapeutic agents for lung cancer.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}