Jove-Journal of Visualized Experiments最新文献

{"title":"In Vivo Calcium Imaging of Taste-Induced Neural Responses in Adult Drosophila.","authors":"Tynan Gacy, Molly Stanley","doi":"10.3791/67917","DOIUrl":"https://doi.org/10.3791/67917","url":null,"abstract":"<p><p>For nearly two decades, in vivo calcium imaging has been an effective method for measuring cellular responses to taste stimuli in the fruit fly model organism, Drosophila melanogaster. A key strength of this methodology is its ability to record taste-induced neural responses in awake animals without the need for anesthesia. This approach employs binary expression systems (e.g., Gal4-UAS) to express the calcium indicator GCaMP in specific neurons of interest. This protocol describes a procedure in which flies expressing GCaMP are mounted with the labellum securely positioned, enabling fluorescence in the brain to be recorded at millisecond resolution under a confocal microscope while a solution is applied to the labellum, stimulating all labellar taste sensilla. The examples provided focus on calcium responses in primary gustatory receptor neurons of D. melanogaster. However, this approach can be adapted to record from other neurons of interest within the brain of Drosophilids or other insect species. This imaging method enables researchers to simultaneously record collective calcium responses from groups of gustatory neurons across the labellum, complementing electrophysiological tip recordings that quantify action potentials from individual neurons. The in vivo calcium imaging technique outlined here has been instrumental in uncovering molecular and cellular mechanisms of chemosensation, identifying unique temporal response patterns in primary taste neurons, investigating mechanisms of gustatory modulation, and exploring taste processing in downstream circuits.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701901","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":"Glycemic Impact on Knee Osteoarthritis Symptoms on Physical, Radiographic, and Inflammatory Markers among Individuals Aged 50 and Over with Diabetes.","authors":"Shi Rui Seow, Sumaiyah Mat, Odelia Hui Hwang Lam, Juliana Fairuz Maktar, Nor Fadilah Rajab, Intan Safinar Ismail, Devinder Kaur Arjit Singh, Suzana Shahar, Maw Pin Tan","doi":"10.3791/67521","DOIUrl":"https://doi.org/10.3791/67521","url":null,"abstract":"<p><p>This study explores hyperglycemia's influence on knee osteoarthritis (KOA) related symptoms, physical performance, physical activity level, radiographic severity, and inflammation in older adults. Prolonged hyperglycemic states contribute to advanced glycation end-product (AGE) formation, which worsens KOA symptoms. Capillary blood glucose (CBG) and glycated hemoglobin A1C (HbA1C) levels are commonly used in laboratory tests for glycemic assessment, offering distinct advantages and limitations. Participants were divided into good and poor glycemic control groups based on their CBG and HbA1C levels. KOA clinical severity and physical activity were measured using the knee injury and osteoarthritis outcome score (KOOS) and international physical activity questionnaire. Physical performance was measured with hand grip strength, gait speed, time-up-and-go (TUG), and 5 times sit-to-stand (5STST). Knee X-rays were performed, and serum enzyme-linked immunosorbent assay (ELISA) analysis was conducted for IL-1β, IL-4, CRP, NF-κB, and AGE. Three hundred recruited participants (mean age [SD] = 66.40 years (5.938) with CBG, of fasting blood sugar > 7.0 mmol/L and random blood sugar > 11.1 mmol/L, (N = 254) were compared with KOOS pain (p=0.008) and symptoms (p=0.017) and 5STST (p=0.015); while HbA1c > 6.3% (N = 93) was compared with 5STST (p=0.002), and AGEs (p=0.022) based on Mann Whitney U test. Logistic regression revealed significant associations between glycemic control and lower limb muscle strength, radiological severity, laboratory markers, and between glycemic status and KOOS pain and symptoms. However, these associations did not remain significant after adjusting for BMI. Poor glycemic status alone was associated with better function in sport and recreation domains after antidiabetic medication adjustment, suggesting anti-inflammatory and analgesic effects that masked the effect of high blood sugar. Future studies could explore the predictive ability of glycemic assessment for poor knee function and physical performance while accounting for the effects of the medication.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701891","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":"Unveiling Xenobiotic Transport and Effects in Isolated Mitochondria: Insights from Respirometric and Enzymatic Assays.","authors":"Luis C Vesga, Jonny E Duque Luna, Stelia C Mendez-Sanchez","doi":"10.3791/67146","DOIUrl":"https://doi.org/10.3791/67146","url":null,"abstract":"<p><p>Mitochondria are often referred to as the cell's powerhouse due to their crucial role in energy production through the electron transport chain (ETC). However, their significance extends far beyond energy production. Dysregulation of mitochondrial bioenergetics can trigger intracellular cascades that impact health and cellular function. Given their physiological importance, there is growing interest in exploring the pharmacological potential of mitochondrial function for developing new therapies and bioproducts. Modulating mitochondrial bioenergetics could provide innovative approaches to address challenges such as neurodegenerative disorders, metabolic diseases, and cancer, as well as to develop bioproducts for controlling pests and vector-borne diseases. This work presents a method for assessing the effects of xenobiotics on the electron transport chain (ETC) using various substrates, including glutamate and NADH for complex I, succinate for complex II, and cytochrome c (both oxidized and reduced) for complexes III and IV. This methodology allows the activation or inhibition of electron transport through mitochondrial complexes to be evaluated using a respirometer and spectrophotometer. The mitochondria can be sourced from isolated mitochondria, fragmented cells, or homogenized tissue from various species. In the laboratory, mitochondrial function has been analyzed in Aedes aegypti and Wistar rats, but this method is also applicable to other species, such as Rhipicephalus microplus and Rhodnius prolixus. This approach provides the basis for theorizing about the existence of uncoupler proteins and species-specific oxidizable substrate preferences influenced by their unique energetic demands. Recent findings offer valuable insights into innovative bioinsecticide design strategies that target mitochondrial function, holding significant potential for effectively controlling vector-borne diseases and pest infestations.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701921","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":"Fixation and Operational Method for Abdominal Massage in T2DM Mice.","authors":"Lizhen Gan, Zhewei Chen, Zhi Zhang, Xinyi He, Xia Wu, Qingbo Wei, Yunchuan Wu","doi":"10.3791/68226","DOIUrl":"https://doi.org/10.3791/68226","url":null,"abstract":"<p><p>Type 2 diabetes mellitus (T2DM) is a rapidly growing global public health issue, affecting over 500 million people worldwide. Although abdominal massage has shown potential benefits in managing T2DM, its effectiveness remains unclear, particularly in animal studies, where challenges such as animal compliance and the need for precise pressure control complicate implementation. To address these challenges, this study introduces a novel abdominal massage simulation device specifically designed for mice. This device provides a practical solution for conducting abdominal massage interventions in a controlled manner while minimizing stress and tissue damage to the animals. It securely restrains the mice's limbs, allowing them to remain conscious during the massage, and offers precise control over both the pressure and frequency of the massage applied to the abdomen. The device's ability to simulate manual abdominal massage with accuracy opens new possibilities for experimental studies assessing its effects on T2DM. The primary goal of this protocol is to investigate the impact of abdominal massage on key T2DM markers such as blood glucose levels, lipid metabolism, and insulin sensitivity in mice. By providing a reliable and reproducible method for abdominal massage, this device can offer valuable insights into its potential as a non-invasive therapeutic intervention for T2DM. The findings from this research may contribute to advancing clinical strategies for diabetes prevention and treatment, particularly in enhancing the understanding of traditional therapies like abdominal massage in modern medical practice.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701798","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":"Identification of Ferroptosis- and Hypoxia-related Genes in iPSC-derived Oligodendrocyte Precursor Cells from Multiple Sclerosis Patients.","authors":"Xia Zhang, Yuming Wang, Ruifang Sui, Yong Zhong, Ping Yi","doi":"10.3791/67055","DOIUrl":"https://doi.org/10.3791/67055","url":null,"abstract":"<p><p>Multiple sclerosis (MS) is a chronic inflammatory disorder characterized by demyelination, with failed remyelination leading to progressive axon loss in chronic stages. Oligodendrocyte precursor cells (OPCs) are critical for remyelination. Recent studies suggest that both hypoxia and ferroptosis play crucial roles in the dysfunctional differentiation of OPCs. This research seeks to identify key genes linked to hypoxia and ferroptosis and immune infiltration characteristics in OPCs derived from induced pluripotent stem cells (iPSCs) of MS patients and to construct a diagnostic model centered on these pivotal genes. We analyzed gene expression data from the GSE196575 and GSE147315 datasets and compared MS patients with healthy individuals. Using weighted gene coexpression network analysis (WGCNA), we pinpointed primary module genes and essential genes associated with hypoxia, ferroptosis, and MS. The ferroptosis Z score and the hypoxia Z score calculated via gene set variation analysis (GSVA) were greater in the iPSC-derived OPCs of MS patients than those of the control group. The implicated genes are predominantly linked to the PI3K/Akt/mTOR pathway, as identified through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A protein-protein interaction (PPI) network of crucial genes revealed 10 central hub genes (COL4A1, COL4A2, ITGB5, ITGB1, ITGB8, ITGAV, VIM, FLNA, VCL, and SPARC). The robust expression of ITGB1, ITGB8, and VIM was validated in the GSE151306 dataset, supporting their role as key hub genes. Additionally, an interaction network between transcription factors (TFs) and hub genes was established via Transcriptional Regulatory Relationships Unraveled by Sentence-based Text (TRRUST), which identified five key TFs. The results of this study could help elucidatenovel biomarkers or therapeutic targets for MS.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701894","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":"Intraoperative Visualization of Subretinal Injection and Retinal Detachment in Rats.","authors":"Alexandre Dentel, Clémence Bradic, Ruben Goulet, Manuel Simonutti, Julie Degardin, Valérie Fradot, Serge Picaud","doi":"10.3791/67483","DOIUrl":"https://doi.org/10.3791/67483","url":null,"abstract":"<p><p>Subretinal injection, the delivery of a solution between the photoreceptors and the retinal pigment epithelium (RPE), creates a subretinal space where components are in direct contact with photoreceptors and RPE cells. This delivery method allows for targeted treatment of these cells. Therapeutics for subretinal injection have been developed and approved, particularly for inherited retinal diseases. In animals, subretinal injection procedures can be challenging due to the lens size, especially in rodents. This article describes methods for subretinal injection in rats, enabling intraoperative visualization and control of both the injection site and the size of the detached area, as performed in humans. The procedure is conducted under general and local anesthesia and requires pupil dilation. Using an ophthalmic microscope, the subretinal injection is performed through a 30 G scleral channel, with the cannula tip gently applied to the retina to create a retinotomy. Volumes ranging from 10-25 µL can be delivered, corresponding to two-fifths to half of the rat retina. Immediate postoperative examinations using fundus photography and optical coherence tomography confirm successful delivery into the subretinal space with visible subretinal fluid. The major risks of this procedure include lens damage (cataract), detachment failure, intravitreal hemorrhage, subretinal hemorrhage, and postoperative keratitis. In addition to delivering therapeutics into the subretinal space, this technique is used to induce short-term or long-term retinal detachment using aqueous or viscous products, respectively. Unlike the transscleral approach, this method enables precise intraoperative positioning of the retinal detachment.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701904","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":"Using 16α-[18F]-Fluoro-17β-Estradiol PET to Visualize Estrogen Receptor α Expression in Human Breast Cancer Xenografts in Female Ovariectomized Mice.","authors":"Sadia Quazi, Nhi Huynh, Angela Rigopoulos, Alexander McDonald, Uwe Ackermann, Andrew Mark Scott, Ingrid Julienne Georgette Burvenich","doi":"10.3791/67151","DOIUrl":"https://doi.org/10.3791/67151","url":null,"abstract":"<p><p>To demonstrate how estrogen receptor alpha (ERα) positive breast cancer xenografts may be visualized in BALB/c nude mice using 16α-[18F]-fluoro-17β-estradiol (¹⁸F-FES) positron emission tomography (PET), ovariectomized BALB/c nude mice were injected with ERα-positive breast cancer cells (MCF-7, 3 × 10⁶ cells; shoulder [n = 10] or 4^th inguinal mammary fat pad [n = 10]) or ERα-negative breast cancer cells (MDA-MB-231, 1 × 10⁶ cells; mammary fat pad [n = 5]). Mice harboring MCF-7 cells received subcutaneous injections of 20 µg of 17β-estradiol (20 µg/20 µL; corn oil:ethanol, 9:1) in the nape of their necks 2 days prior to cell injection, followed by daily injections five times per week for 5 weeks. Tumor volumes were measured according to the formula: (L*W²)/2 (L; length, W; width). Once tumor volumes reached approximately 100 mm³, 17β-estradiol injections were halted 2 days prior to mice receiving ¹⁸F-FES for PET imaging to avoid competitive binding with ERα. Upon ¹⁸F-FES administration via the lateral tail vein, PET/MRI was performed for 15 min at 1 h to 1.5 h post-injection. ¹⁸F-FES uptake was not observed in ERα-negative, MDA-MB-231 tumor-bearing mice. ¹⁸F-FES uptake was most pronounced in mice harboring MCF-7 tumors in the shoulder. In MCF-7 tumors grown in the inguinal mammary fat pad, ¹⁸F-FES uptake was less visible, as the intestinal excretion pattern of ¹⁸F-FES obscured the radioactivity detectable in these tumors. To use ¹⁸F-FES PET as a tool to visualize ERα expression in ERα-positive breast xenografts, we demonstrate that the visibility of ¹⁸F-FES uptake is clear in tumors located away from the abdominal region of mice, such as in the shoulder.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 217","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701927","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":"Non-Invasive Visualization of Nailbed Microvascular Morphology in Mice Using Capillaroscopy.","authors":"Olivia L Bossardet, Clara C Cousins, Joseph M Holden, Vincent Yao, Kristin L Clark, Louis R Pasquale, Emmanuel S Buys, Lauren K Wareham","doi":"10.3791/67529","DOIUrl":"https://doi.org/10.3791/67529","url":null,"abstract":"<p><p>Imaging microcapillary networks of the skin in humans using nailfold capillaroscopy (NFC) has underscored the importance of microcirculation as a target organ system in critical systemic illnesses. Nailfold capillaroscopy is applied clinically to detect peripheral microvascular dysfunction and abnormalities in a range of systemic conditions, including rheumatic, cardiac, ocular (e.g., glaucoma), and endocrine disorders (e.g., hypertension and diabetes mellitus). NFC is useful not only in detecting peripheral systemic microvasculature disruption but also in assessing drug efficacy. However, translating clinical NFC findings to animal disease models can be challenging. Detecting microvascular dysfunction or abnormalities in animals is often invasive (e.g., endoscopic), carried out ex vivo (e.g., post-mortem imaging of tissues), or expensive, requiring specialized equipment such as those used in microcomputed tomography and photoacoustic imaging techniques. Developing quick, non-invasive, and inexpensive techniques to image peripheral microvasculature in animal models of disease is warranted to decrease research expenses and increase translatability to the clinic. Capillaroscopy has previously been used to visualize the nailfold microvasculature in animal models, including in guinea pigs and mice, thus demonstrating the capability of capillaroscopy as a non-invasive imaging tool in animal models. This study provides a protocol that applies capillaroscopy to a mouse nailbed, allowing researchers to easily and inexpensively assess the morphology of its microvasculature. Representative images of typical nailbed microvascular architecture in wild-type mice using two commonly used laboratory strains, SV129/S6 and C57/B6J, are provided. Further studies using this method are essential for applying nailbed capillaroscopy to a wide range of mouse disease models with peripheral microvascular abnormalities.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 216","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143651582","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":"In vivo and In vitro Infection of Potato Roots with Plant Parasitic Nematodes for the Assessment of Induced Structural Changes.","authors":"Jorge M S Faria, Pedro Barbosa, A Cristina Figueiredo, Manuel Mota, Cláudia S L Vicente","doi":"10.3791/67756","DOIUrl":"https://doi.org/10.3791/67756","url":null,"abstract":"<p><p>Soil-dwelling plant parasitic nematodes (PPNs) are important potato pests that cause lesions and/or change plant roots structure, leading to reduced crop fitness and productivity. Research on the cellular and subcellular mechanisms of PPNs infection and development can resort to field plants or seedlings under greenhouse conditions. Field studies are more representative of natural environments but are subjected to the unpredictability of environmental conditions that can heavily influence research outcomes. Greenhouse studies allow higher control over environmental variables and higher safety against contaminants or pathogens. However, in some hosts, genetic diversity becomes an important factor of variability and influences the host-parasite complex response. We have developed in vitro co-cultures of transgenic roots with PPNs as a reliable alternative that occupies less space, requires less time to obtain, and is free from contamination or from host genetic variability. Co-cultures are obtained by introducing aseptic PPNs to host in vitro transgenic roots. They can be maintained indefinitely, which makes them excellent support for keeping collections of reference PPNs. In the present work, a protocol is detailed for the controlled infection of in vivo potato roots with the root lesion nematode and for establishing in vitro co-cultures of potato transgenic roots with the root-knot nematode. The in vitro co-cultures provided a laboratory proxy for the natural potato infection condition and produced nematode life stages irrespective of season or climate conditions. Additionally, the methodology used for structural analysis is detailed using histochemistry and optical microscopy. The acid fuchsin dye is used to follow nematode attack sites on roots, while differential staining with Periodic acid-Schiff (PAS) and toluidine blue O highlights nematode structures in potato internal root tissue.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 216","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143651501","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":"Cryosectioning and Immunostaining of Mouse Retina.","authors":"Yongqiong Lin, Yingjie Tong, Tongdan Zou, Jiajia Wang, Houbin Zhang","doi":"10.3791/67622","DOIUrl":"https://doi.org/10.3791/67622","url":null,"abstract":"<p><p>Tissue sectioning and immunohistochemistry are essential techniques in histological and pathological studies of retinal diseases using animal models. These methods enable detailed examinations of tissue morphologies and the localization of specific proteins within the tissue, which provide valuable insights into disease processes and mechanisms. Mice are the most widely used model for this purpose. However, because mouse eyeballs are small and mouse retinas are extremely delicate tissues, obtaining high-quality retinal sections and immunostaining images from mouse eyeballs is typically challenging. This study describes an improved protocol for cryosectioning mouse retinas and performing immunohistochemistry. An essential point of this protocol involves coating the eyeball with a layer of super glue, which prevents deformation of the eyeballs during the processes of cornea removal, lens extraction, and embedding. This step ensures the integrity of retinal morphologies is well preserved. This protocol highlights critical technical considerations and optimization strategies for consistently producing high-quality retinal sections and achieving excellent immunostaining results.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 216","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143651479","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}