JoVE Neuroscience is a multidisciplinary section devoted to investigations of the structure, function, physiology, and pathophysiology of the brain and nervous system. Included methodologies range from molecular and cellular level studies to full central and peripheral neural systems. Potential treatment platforms and surgical techniques for neurological diseases and disorders are also presented in this section.

Neuroscience

JoVE Medicine serves as a connection between basic research and clinical applications. Articles may present medical procedures, case studies or clinical trial methodologies. Included studies are often modeled towards predictive, preventative, personalized, and practical approaches to treatment of disease. The goal of research methodologies in this section is to improve overall patient care and prognosis.

Medicine

JoVE Immunology and Infection includes content from a variety of fields working toward a shared initiative of improving global health. Techniques evaluating the biological response to pathogens from the molecular to organismal level are presented as well as therapeutic agents and their efficacy in treating disease.

Immunology and Infection

JoVE Genetics publishes video articles demonstrating experimental approaches to investigate gene function and regulation. This includes methods to study gene expression, mutagenesis, epigenetics, genetic determinants of development, genetic disorders, population genetics and evolution.

Genetics

JoVE Environment is dedicated to research methodologies that address environmental concerns and seek to better understand Earth's ecosystem.  Special consideration is given to experimental methodologies that assess society’s impact on the environment, suggest solutions for protecting Earth’s resources, and develop sustainable fuel sources. Moving towards an environmentally conscious planet is a collaborative effort with contributions from both life and physical scientists.

Environment

JoVE Engineering encompasses a broad range of experimental and instrumental techniques utilized in physics research. Investigations in this area strive to address and answer a broad range of scientific questions, such as device mechanisms and efficiencies, using physical tools. This approach often requires a combination of specialties, and research in this area tends to be interdisciplinary with contributions from mechanical, electrical, and chemical engineers.

Engineering

JoVE Developmental Biology publishes research methodologies in the field of developmental biology. This includes methods that are used to study biological development in vitro and in vivo at the molecular, cellular, tissue, organ and whole organism levels.

Developmental Biology

JoVE Chemistry surveys a broad scope of chemical disciplines. This collection includes fundamental and applied research that investigates molecular interactions, properties, and structure spanning in the following core divisions: analytical, organic, organometallic, inorganic, bioinorganic, and biochemistry. This section also includes the design and preparation of materials and small molecules for advanced or unique applications.

Chemistry

JoVE Cancer Research illustrates experimental approaches in biomedical research and clinical practice aimed at understanding, detecting, treating and preventing cancer. The video articles in this section demonstrate methods to study the cellular and molecular mechanisms of cancer, animal models of cancer research, development of new therapies and diagnostics, tumor imaging, chemical and radiation treatments, surgical procedures and improvements in therapy. These video articles offer researchers and clinical practitioners an efficient way to learn the technologies and experimental methods in cancer research.

Cancer Research

JoVE Biology welcomes all general biology research methodologies. Content in this section canvases all fields of cell, molecular, and organismal biology, ranging from new applications of standard techniques to novel approaches aimed at understanding the functions of life and living organisms. This diverse section includes, but is not limited to, techniques in physical biology, cellular biochemistry, genetics, physiology, systems biology and a combination of eukaryotic and prokaryotic model systems.

Biology

JoVE Bioengineering merges both physical and life sciences to understand and predict biological processes. Applying physical science tools to life science questions allow for the discovery of better technologies to measure, diagnose, and clinically treat disease.

Bioengineering

JoVE Biochemistry publishes experiments that investigate the structure, function, and interactions of biological molecules. The video articles in this section demonstrate methods to study nucleic acids, proteins, carbohydrates and lipids as well as enzyme function, protein folding, molecular interactions and biochemical reactions.

Biochemistry

JoVE Behavior documents observational and experimental techniques that seek to understand human and animal behavior. Work in this field includes analysis of genetic causes and physiological responses to internal or external stimuli as well as the implicated brain regions.

Behavior

Arthropod borne viruses (arboviruses) are a group of mosquito-borne pathogens that are globally distributed. Since the 2016 outbreak of Zika virus (ZIKV) in the Americas that brought these infections to public awareness, arboviruses continue to impose a considerable public health burden. Their emergence in previously non-endemic regions is a cause of public health concern. The surveillance and control of arboviral infections is a current priority for the World Health Organization (WHO), which launched a global strategic plan to fight arboviral diseases in 2024. The multiplex&#174; technology has revolutionized the field of serological surveillance, allowing simultaneous measurement of multiple antibodies while requiring minimum sample volume. Building on expertise in the development of multiplex serological tests, a bead-based multiplex assay for the detection of antibodies to arbovirus antigens was developed. The assay comprises multiple antigens specific to common arboviruses, including dengue (DENV), yellow fever (YFV), Zika, and chikungunya (CHIKV) viruses. A semi-automated method was used to covalently bind antigens to microsphere beads. This new protocol is highly effective, enabling the coupling of up to 96 antigens in a single experiment while ensuring reproducible results. Median Fluorescence Intensity (MFI) data were acquired using a user-friendly fluorescence imaging system.When applied to human serum samples from endemic countries, we anticipate that this assay will constitute a powerful tool for sero-surveillance efforts.

a-multiplex-serological-assay-for-detection-antibody-responses-to

A Multiplex Serological Assay for the Detection of Antibody Responses to Arboviruses

Arthropod borne viruses (arboviruses) are a group of mosquito-borne pathogens that are globally distributed. Since the 2016 outbreak of Zika virus (ZIKV) ...

Anti-MDA5 autoantibodies are critical biomarkers for dermatomyositis (DM), amyopathic dermatomyositis (CADM), and polymyositis (PM), particularly in identifying patients at risk of rapidly progressive interstitial lung disease (RP-ILD). Early detection of these autoantibodies is essential to improve patient outcomes, as delayed diagnosis often leads to poor prognoses. Currently, radioimmunoassay is the gold standard for detecting anti-MDA5, but its use is limited by high costs, lengthy procedures, and the need for specialized expertise. Additionally, the blot test, a widely used clinical tool, exhibits a high false-positive rate for MDA5 autoantibodies, potentially compromising diagnostic accuracy. To address these limitations, we propose a non-radioactive, highly sensitive, and standardized confirmatory testing method using Immunocytochemistry (ICC). This protocol involves treating HeLa cells with an MDA5 construct, permeabilizing the cells with Triton X-100 to facilitate binding of primary anti-MDA5 autoantibodies, and detecting bound antibodies using enzyme-conjugated secondary antibodies (e.g., horseradish peroxidase) with DAB chromogen for microscopy imaging. ICC offers a practical, cost-effective, and high-sensitivity approach for visualizing anti-MDA5 autoantibodies within cellular structures. By integrating ICC as a supplementary confirmatory procedure, this study aims to enhance the reliability of anti-MDA5 detection, thereby improving diagnostic and prognostic strategies for RP-ILD in DM, CADM, and PM patients.

detection-anti-mda5-autoantibodies-using-hela-cells

Detection of Anti-MDA5 Autoantibodies Using HeLa Cells and Immunocytochemistry with Light Microscopy

Anti-MDA5 autoantibodies are critical biomarkers for dermatomyositis (DM), amyopathic dermatomyositis (CADM), and polymyositis (PM), particularly in ...

Chronic pancreatitis (CP) is a severe inflammatory condition that can destroy the pancreas, lead to diabetes, and increase pancreatic cancer risk, severely impacting patients' quality of life. Addressing CP is a critical research focus in gastroenterology. We developed a new animal model to overcome the limitations of existing ones, using 8-week-old C57BL/6 male mice.
After exposing the pancreas through abdominal surgery on anesthetized mice, we clamped the common bile duct near the liver to prevent methylene blue from entering it. A capillary was inserted into the bile duct to inject the dye, and after the pancreatic duct was stained. After the pancreatic duct had been visualized, the pancreas was selected for electrocoagulation in the middle and lower 2/3 of the position between the common bile duct and the superior pancreaticoduodenal artery. Electrocoagulation was terminated when the blue staining solution could not be recognized. The abdomen was then closed, and the mice were normally raised. At 7, 14, 21, and 28 days post surgery, the pancreas were examined using HE staining, Masson staining, and immunohistochemistry, and blood serum was analyzed for biochemical parameters. The pathology after 14 days closely matched the characteristics of chronic pancreatitis, as did the serum biochemical parameters.

an-obstructive-chronic-pancreatitis-model-established-through

An Obstructive Chronic Pancreatitis Model Established Through Electrocoagulation

Chronic pancreatitis (CP) is a severe inflammatory condition that can destroy the pancreas, lead to diabetes, and increase pancreatic cancer risk, ...

Ticks are blood-feeding ectoparasites recognised for their ability to transmit several infectious pathogens to humans and animals. Tick-borne pathogen (TBP) surveillance generally relies on detecting pathogens in homogenates of entire ticks, including those acquired during blood feeding, which may not be transmitted by the tick. To better understand the pathogen transmission mechanism, it is essential to investigate the dissemination and concentration of TBPs in various tick tissues, which is important for elucidating transmission mechanisms in ticks and determining their vector competence. This, in turn, rests on procedures for isolating saliva, haemolymph, salivary glands, and the midgut of individual ticks in order to examine the location of TBPs within these tissues and fluids. In this study, we describe a protocol for tick tissue collection, including a novel procedure for the collection of haemolymph. The results confirm that Rickettsia africae occurs at higher levels in the haemolymph of Amblyomma gemma, affirming the likely role of this tick species as a competent vector for R. africae. In contrast, Hyalomma dromedarii exhibited high rates of R. africae in the midgut, but the bacterium was absent in the haemolymph. The presence of TBPs in the haemolymph is therefore a valuable indicator of vector competence, and rests on the availability of a method that ensures ease of collection.

extraction-saliva-haemolymph-salivary-glands-midgut-from-individual

Extraction of Saliva, Haemolymph, Salivary Glands, and Midgut from Individual Ticks (Acari: Ixodidae)

Ticks are blood-feeding ectoparasites recognised for their ability to transmit several infectious pathogens to humans and animals. Tick-borne pathogen ...

Highly multiplexed imaging enables the study of functionally diverse cells and their niches within their native tissue environments. Iterative Bleaching Extends Multiplexity (IBEX) is a cyclic immunolabeling and fluorophore inactivation technique that allows for multiple markers to be visualized on the same tissue section. Captured images can be subsequently analyzed to acquire single-cell data to define cell clusters, their localizations, and neighboring cell types. Interpreting these data relies on the ability to distinguish true marker expressions from sources of background inherent to fluorescence microscopy. Spectral IBEX, an adaptation of the IBEX protocol, integrates spectral confocal detection with computational unmixing and incorporates heparin blocking to reduce charge-based off-target binding. This combination improves the signal-to-background ratio, suppresses tissue autofluorescence, and minimizes bleed-through while also reducing acquisition time compared to conventional multi-track confocal imaging. Application to human nasal polyp tissue, a model characterized by high eosinophil content and strong autofluorescence, demonstrated reliable imaging of 26 markers across structural, immune, and cell state compartments over six imaging rounds. The resulting workflow generates high-dimensional, spatially resolved proteomic information that captures complex tissue architecture and cellular niches. Together, this optimized approach provides a robust and broadly applicable strategy for multiplexed imaging, particularly suited to tissues where autofluorescence and non-specific staining limit conventional approaches.

high-plex-imaging-using-spectral-confocal-microscopy-to-minimize-non

High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence

Highly multiplexed imaging enables the study of functionally diverse cells and their niches within their native tissue environments. Iterative Bleaching ...

Tuberculosis (TB), primarily a pulmonary disease, manifests extrapulmonary involvement in 20% of cases, though concurrent infections in distant sites like the hand and abdomen are exceedingly rare. This report presents an exceptionally rare case of concurrent tuberculosis osteomyelitis of the hand and peritoneal tuberculosis in a 19-year-old male-an unusual dual presentation not commonly documented in the literature. The patient had an 8-month history of progressive right little finger swelling, initially misdiagnosed as an enchondroma, and subsequently developed acute abdominal pain and ascites, with computed tomography (CT) revealing diffuse peritoneal thickening and peritonitis. Ascitic analysis showed exudative fluid with elevated CA-125, and an interferon-gamma release assay (IGRA) was positive, prompting TB suspicion. Histopathology of debrided hand tissue revealed granulomatous inflammation with necrosis, and Mycobacterium tuberculosis was confirmed by acid-fast staining and fluorescence in situ hybridization (FISH). Treatment combined surgical debridement, bone grafting, and vacuum-assisted drainage for the hand lesion with a 12-month HRZE regimen. Abdominal TB resolved with medical therapy alone. The patient achieved complete recovery with restored finger function and no recurrence over 2 years. This case underscores TB's protean manifestations, emphasizing diagnostic challenges in paucibacillary disease and the utility of IGRA and molecular confirmation (FISH). Clinicians should maintain high suspicion for TB in atypical osteolytic lesions or idiopathic serositis, particularly in endemic regions, to ensure timely multidisciplinary management.

an-unusual-case-of-tuberculosis-infection-of-the-hand-with-peritonitis

An Unusual Case of Tuberculosis Infection of the Hand with Peritonitis

Tuberculosis (TB), primarily a pulmonary disease, manifests extrapulmonary involvement in 20% of cases, though concurrent infections in distant sites like ...

Adoptive T cell therapies, including chimeric antigen receptor (CAR) T cells, have demonstrated impressive clinical activity against hematologic malignancies and are showing promise in treating solid tumor malignancies. Despite these successes, multiple mechanisms of resistance to T cell immunotherapy have been identified that limit therapeutic success, including loss of or weak expression of target antigens, development of T cell exhaustion, and the presence of an immunosuppressive tumor microenvironment. Multiple cell engineering strategies have been developed to overcome these mechanisms of resistance. However, multiple mechanisms of resistance can occur simultaneously, necessitating a combination of multiple engineering strategies to optimize anti-tumor activity. Vector packaging constraints limit the delivery of large amounts of genetic information to T cells and present a challenge in co-expressing multiple engineered constructs. Here, we describe a protocol for co-transducing T cells with two vectors to encode multiple transgenes, thereby increasing the number of engineered functions. By co-expressing a leucine zipper-based cell sorting methodology, termed Zip-sort, we direct selective immunomagnetic purification of dual-transduced cells that have incorporated two distinct vectors.

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Leucine Zipper-based Cell Sorting for Purification of Dual-vector-transduced Cells

Adoptive T cell therapies, including chimeric antigen receptor (CAR) T cells, have demonstrated impressive clinical activity against hematologic ...

The emergence of multidrug-resistant tuberculosis (MDR-TB) necessitates rapid and accurate drug susceptibility testing (DST) methods to guide effective treatment. This study introduces a methodology combining the resazurin microtiter assay (REMA) and checkerboard assay to determine minimum inhibitory concentration (MIC) and evaluate drug-drug interactions of anti-tuberculosis drugs against Mycobacterium tuberculosis. The REMA, adapted to a 96-well format, leverages the reduction of resazurin dye by metabolically active M. tuberculosis as a visual indicator of drug susceptibility. Varying concentrations of anti-TB drugs are tested against M. tuberculosis isolates, and color changes are observed to determine the MIC. Subsequently, a checkerboard assay is employed to assess potential synergistic, additive, or antagonistic effects between drug combinations. This simple and inexpensive method yields results within seven days, offering a significant advantage over traditional DST methods. This method provides valuable insights into the DST of M. tuberculosis isolates and facilitates the identification of promising drug combinations for improved treatment outcomes against MDR-TB.

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Detection of Drug Susceptibility and Drug Interactions in Mycobacterium tuberculosis using Resazurin Microtiter and Checkerboard Assay

The emergence of multidrug-resistant tuberculosis (MDR-TB) necessitates rapid and accurate drug susceptibility testing (DST) methods to guide effective ...

Crohn disease (CD) is a chronic, relapsing inflammatory condition characterized by segmental, transmural inflammation that can affect any part of the gastrointestinal tract. A hallmark of CD is impaired intestinal barrier function, and intestinal fibrosis is a common complication. SHIP-deficient (SHIP-/-) mice spontaneously develop gut barrier dysfunction, ileal inflammation, and fibrotic pathology, recapitulating key features of CD-like ileitis. The SHIP-/- mice serve as a valuable model for testing anti-inflammatory and anti-fibrotic therapies in CD.
Here, we describe methods to evaluate therapeutic interventions in this model, using dexamethasone as an example of an effective therapy. We provide a step-by-step protocol for characterizing disease and treatment response, including in vivo assessment of gut permeability measured by FITC-dextran, as well as gross pathological examination and detailed histological analyses. Histological assessments incorporate hematoxylin and eosin (H&#38;E) staining for inflammation, Masson&#39;s trichrome staining for fibrosis, and Alcian blue/Periodic Acid-Schiff (PAS) staining for goblet cell hyperplasia and hypertrophy. Additionally, cytokines and inflammatory markers in ileal tissue are quantified to assess the inflammatory state. Together, these methods provide a comprehensive framework for evaluating treatment efficacy in the SHIP-/- mouse model of CD-like ileitis. This protocol is broadly applicable to investigators studying gut inflammation, mucosal healing, and intestinal fibrosis.

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Evaluating Therapeutic Interventions in the SHIP-deficient Mouse Model of Crohn Disease-like Ileitis and Fibrosis

Crohn disease (CD) is a chronic, relapsing inflammatory condition characterized by segmental, transmural inflammation that can affect any part of the ...

Preterm birth continues to be a primary cause of neonatal mortality and morbidity globally, with limited preventative therapies. There is an imperative need for clinically 
relevant and reproducible animal models for the translation of innovative interventions 
to become a reality. Various mouse models of preterm birth exist. However, they do not recapitulate human neonatal morbidities, including neuropathological changes and 
lung injury, which are characteristic sequelae of ascending infection and preterm birth. We have characterized a model which, to our knowledge, is the only one that mimics the human scenario whereby these neonatal morbidities are&#160;present following the induction of preterm birth by a live bacterium that ascends from the vagina. This paper demonstrates the&#160;intravaginal administration of a bioluminescent strain of Escherichia coli and details the assessment of neuropathological outcomes and lung injury using immunohistochemistry and histology techniques.

modeling-ascending-vaginal-infection-preterm-birth-neonatal-morbidity

Modeling Ascending Vaginal Infection, Preterm Birth, and Neonatal Morbidity in Mice

Preterm birth continues to be a primary cause of neonatal mortality and morbidity globally, with limited preventative therapies. There is an imperative ...

immunology and infection

immunology and infection