Animals, cell culture and transfection, RT-PCR, SPHK activity assay, S1P measurement, confocal imaging, endothelial permeability determination, and lung vascular permeability determination are described in the expanded Material and Methods section in the online data supplement, available at http://circres.ahajournals.org .
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Vascular Permeability
Vascular Permeability
Vascular permeability refers to the ability of substances, such as fluids, macromolecules, and cells, to move across the endothelial barrier of blood vessels.
This process is essential for the exchange of nutrients, gases, and waste products between the bloodstream and surrounding tissues.
Alterations in vascular permeability can contribute to the development of various pathological conditions, including inflammation, edema, and tumor angiogenesis.
Understanding and optimizing the measurement of vascular permeability is crucial for research in fields such as cardiovascular biology, oncology, and drug delivery.
PubCompare.ai's AI-driven protocol comparison tool can help streamline your vascular permeability research by allowing you to easily locate, evaluate, and compare protocols from literature, pre-prints, and patents, ensuring your studies are grounded in the best available evidence and leveraging AI-powered analysis to identify the most accurate and reproducible methods.
This process is essential for the exchange of nutrients, gases, and waste products between the bloodstream and surrounding tissues.
Alterations in vascular permeability can contribute to the development of various pathological conditions, including inflammation, edema, and tumor angiogenesis.
Understanding and optimizing the measurement of vascular permeability is crucial for research in fields such as cardiovascular biology, oncology, and drug delivery.
PubCompare.ai's AI-driven protocol comparison tool can help streamline your vascular permeability research by allowing you to easily locate, evaluate, and compare protocols from literature, pre-prints, and patents, ensuring your studies are grounded in the best available evidence and leveraging AI-powered analysis to identify the most accurate and reproducible methods.
Most cited protocols related to «Vascular Permeability»
Animals
Biological Assay
Cell Culture Techniques
Dietary Supplements
Endothelium
Lung
Permeability
Reverse Transcriptase Polymerase Chain Reaction
Transfection
Vascular Permeability
Details of the mathematical model can be found in the appendix. Briefly, a Krogh cylinder geometry was used where a cylindrical blood vessel segment is surrounded by tissue with a radius approximately equal to half the local inter-capillary distance (Figure 1B ). Blood flows from the arterial end, where a systemic two-compartment model with biexponential decay defines the concentration. The local blood concentration is determined by the blood velocity, permeability of the vessel wall, and fraction of free drug (not bound to blood cells or plasma proteins). Cellular uptake in the blood was ignored (File S1 ) given the slower kinetics relative to blood flow [22] (link). A mixed boundary condition is used at the capillary interface, where the flux at the capillary wall determined by the permeability is equal to the diffusive flux into the tissue. In the tissue, the free drug undergoes radial and axial diffusion along with agent specific reaction terms. For small molecules, this involves cellular uptake and metabolism (e.g. oxygen utilization, irreversible trapping over short time scales by FDG phosphorylation, reversible uptake for doxorubicin). For antibodies, this involves reversible binding and dissociation with irreversible internalization. Due to the lack of functional lymphatics in tumors, lymphatic drainage was ignored [23] (link). The following equations defined the plasma concentration, plasma tissue interface, and tissue concentration (for first order kinetics):
where [C]plasma is the total concentration of drug in the plasma, t is time, v is the local blood velocity, L is the length along the vessel segment, Rcap is the capillary radius, H is the hematocrit, P is the vessel wall permeability, ffree is the fraction of drug that is unbound, [C]tissue,free is the unbound concentration in the tissue (overall/pseudohomogenous concentration), and epsilon the void fraction. D is the effective diffusion coefficient in tissue, r is the radial distance from a vessel, and krxn defines the local reaction rate (which is first order in this example equation).
The method of lines was used with axial and radial variations and solved with a stiff solver using Matlab (The Mathworks; Natick, MA). A sparse Jacobian was defined to decrease simulation times.
where [C]plasma is the total concentration of drug in the plasma, t is time, v is the local blood velocity, L is the length along the vessel segment, Rcap is the capillary radius, H is the hematocrit, P is the vessel wall permeability, ffree is the fraction of drug that is unbound, [C]tissue,free is the unbound concentration in the tissue (overall/pseudohomogenous concentration), and epsilon the void fraction. D is the effective diffusion coefficient in tissue, r is the radial distance from a vessel, and krxn defines the local reaction rate (which is first order in this example equation).
The method of lines was used with axial and radial variations and solved with a stiff solver using Matlab (The Mathworks; Natick, MA). A sparse Jacobian was defined to decrease simulation times.
Antibodies
Arteries
BLOOD
Blood Cells
Blood Circulation
Blood Vessel
Capillaries
Cells
Diffusion
Doxorubicin
Drainage
Kinetics
Lymphatic System
Metabolism
Neoplasms
Oxygen
Permeability
Pharmaceutical Preparations
Phosphorylation
Plasma
Plasma Proteins
Radius
Tissues
Urination
Vascular Permeability
Volumes, Packed Erythrocyte
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Arteries
Blood Vessel
Contrast Media
Diffusion
Extracellular Space
gadobutrol
Gadolinium
Gadovist
Microtubule-Associated Proteins
Necrosis
Neoplasms
Normal Saline
Perfusion
Plasma
Radionuclide Imaging
Tissues
Vascular Permeability
EB intravenous injection is a well-established method for measuring lung vascular permeability. We compared this new method with the EBA assay after intravenous injection of EB. Eighteen mice were divided into three groups, which included the control and 6 h and 24 h model groups. The mice of the model groups were instilled i.n. with 4 mg of LPS/kg b.w., and the mice of the control group were instilled i.n. with 50 µl of PBS. One hour before the mice were sacrificed, 10 mg of FITC-Dextran/kg b.w. was instilled i.n. into the airway of the model and control groups. The FI of the plasma FITC-Dextran was measured as described above.
EB (20 mg/kg b.w.; Sigma-Aldrich, St Louis, MO, USA) was injected into the retroorbital venous sinus in the model and control mice for 30 minutes as previously reported [6] (link), [10] (link) before all of the mice were euthanized. The lungs were perfused free of blood (perfusion pressure of 5 mmHg) with PBS containing 2 mM EDTA via thoracotomy. The right lung was homogenized in PBS (1 ml/100 µg tissue), incubated with 2 volumes of formamide (18 hours at 60°C), and centrifuged at 5,000×g for 30 minutes. The optical density of the supernatant was determined spectrophotometrically at 620 nm and 740 nm using a Synergy H1 plate reader (BioTek). The extravasated EBA concentration in the lung homogenate was calculated against a standard curve (micrograms of Evans Blue dye per lung). The following formula was used to correct the optical densities for contamination with heme pigments: E620(corrected) = E620(raw)−(1.426×E740(raw)+0.030). Given the different routes of administration, the fold changes relative to the basal and not the absolute values were compared.
EB (20 mg/kg b.w.; Sigma-Aldrich, St Louis, MO, USA) was injected into the retroorbital venous sinus in the model and control mice for 30 minutes as previously reported [6] (link), [10] (link) before all of the mice were euthanized. The lungs were perfused free of blood (perfusion pressure of 5 mmHg) with PBS containing 2 mM EDTA via thoracotomy. The right lung was homogenized in PBS (1 ml/100 µg tissue), incubated with 2 volumes of formamide (18 hours at 60°C), and centrifuged at 5,000×g for 30 minutes. The optical density of the supernatant was determined spectrophotometrically at 620 nm and 740 nm using a Synergy H1 plate reader (BioTek). The extravasated EBA concentration in the lung homogenate was calculated against a standard curve (micrograms of Evans Blue dye per lung). The following formula was used to correct the optical densities for contamination with heme pigments: E620(corrected) = E620(raw)−(1.426×E740(raw)+0.030). Given the different routes of administration, the fold changes relative to the basal and not the absolute values were compared.
Biological Assay
BLOOD
Edetic Acid
Evans Blue
fluorescein isothiocyanate dextran
formamide
Heme
Lung
Mus
Perfusion
Pigmentation
Plasma
Pressure
Sinuses, Nasal
Thoracotomy
Tissues
Vascular Permeability
Veins
Vision
Biopsy
Blood Vessel
Capillaries
Capillary Leak Syndrome
Chinese
Patients
Skin
Specimen Collection
Vascular Permeability
Vision
Most recents protocols related to «Vascular Permeability»
Example 7
The efficacy of UBX1967 was studied in a mouse model of diabetic retinopathy, by a single administration of streptozotocin (STZ).
C57BL/6J mice of 6- to 7-week were weighted and their baseline glycemia was measured (Accu-Chek, Roche). Mice were injected intraperitoneally with STZ (Sigma-Alderich, St. Lois, Mo.) for 5 consecutive days at 55 mg/Kg. Age-matched controls were injected with buffer only. Glycemia was measured again a week after the last STZ injection and mice were considered diabetic if their non-fasted glycemia was higher than 17 mM (300 mg/dL). STZ treated diabetic C57BL/6J mice were intravitreally injected with 1l of UBX1967 (2 μM or 20 μM, formulated as a suspension in 0.015% polysorbate-80, 0.2% Sodium Phosphate, 0.75% Sodium Chloride, pH 7.2) at 8 and 9 weeks after STZ administration. Retinal Evans blue permeation assay was performed at 10 weeks after STZ treatment.
Animal Model
Biological Assay
Blood Vessel
Buffers
Choroid
Diabetes Mellitus
Diabetic Retinopathy
Evans Blue
Figs
Mice, Inbred C57BL
Mus
Polysorbate 80
Retina
Retinal Diseases
Sodium Chloride
sodium phosphate
Streptozocin
Vascular Permeability
The positional candidate genes were ranked based on their predicted association with seven functional terms related to the Bphs phenotype: Cardiac, G-protein coupled receptor, Histamine, Pertussis toxin, Type I hypersensitivity, Vascular Permeability, and ER/EMC/ERAD. Gene Weaver74 (link) was used to identify genes annotated with each term. Each term was entered the Gene Weaver homepage (https://geneweaver.org ) and search restricted to human, rat, and mouse genes, and to curated lists only. Mouse homologs for each gene were retrieved using the batch query tool in MGI (http://www.informatics.jax.org/batch_data.shtml ). In addition, using the Gene Expression Omnibus (GEO) and PubMed, additional gene expression data sets were retrieved for each phenotype term. Final gene lists consisted of the unique set of genes associated with each process term.
Benign Prostatic Hyperplasia
G-Protein-Coupled Receptors
Gene Expression
Genes
Heart
Histamine
Homo sapiens
Immediate Hypersensitivity
Mice, Laboratory
Pertussis Toxin
Phenotype
Vascular Permeability
For the permeability assay of keratinocyte monolayers, we used an in vitro vascular permeability assay kit (Sigma-Aldrich). The N/TERT were plated on the 18 mm coverslips prepared according to the manufacturer’s protocol and the experimental treatment was performed as described above. Then, the samples were treated with Fluorescein-Streptavidin for 5 min and fixed with 4% paraformaldehyde for 15 min, washed in PBS 3 times, mounted on slides and imaged with the confocal microscope. The areas inside large keratinocyte islands were chosen for the imaging.
To test the basal-to-apical permeability of RHE cultures, we adapted a method used in.104 (link) The RHEs were cultured and stimulated as described above. After 24 h of stimulation, the biotin stock solution (10 mg/mL in water, Tocris, Cat.# 7302) was added directly to the media at the bottom of the inserts to the final concentration of 0.5 mg/mL. The cultures were incubated for 1 h. Then RHEs were fixed in 4% formaldehyde overnight and embedded in paraffin. 5 μm sections were cut, deparaffinized and directly stained with Fluorescein-Streptavidin (Sigma-Aldrich, Cat.# 17–10398) at 1:2000 30 min in PBS.
For the apical-to-basal permeability assay on RHE cultures, RHE cultures were moved from deep well plates to standard 12 well plates after 11 days of airlift (10 days of standard airlifted culture followed by 24 h of stimulation). In 12 well plate, 1.850 mL medium with continuous respective stimulation was added to the bottom well and 0.6 mL medium with continuous respective stimulation and 0.2 mM Na-Fluorescein (Sigma-Aldrich) was placed in the upper chamber, on top of the cornified layer. The fluorescence intensity in the bottom well was measured after 24 h of incubation with fluorescein (meas. filter 492 nm, ref. filter 570 nm).
To test the basal-to-apical permeability of RHE cultures, we adapted a method used in.104 (link) The RHEs were cultured and stimulated as described above. After 24 h of stimulation, the biotin stock solution (10 mg/mL in water, Tocris, Cat.# 7302) was added directly to the media at the bottom of the inserts to the final concentration of 0.5 mg/mL. The cultures were incubated for 1 h. Then RHEs were fixed in 4% formaldehyde overnight and embedded in paraffin. 5 μm sections were cut, deparaffinized and directly stained with Fluorescein-Streptavidin (Sigma-Aldrich, Cat.# 17–10398) at 1:2000 30 min in PBS.
For the apical-to-basal permeability assay on RHE cultures, RHE cultures were moved from deep well plates to standard 12 well plates after 11 days of airlift (10 days of standard airlifted culture followed by 24 h of stimulation). In 12 well plate, 1.850 mL medium with continuous respective stimulation was added to the bottom well and 0.6 mL medium with continuous respective stimulation and 0.2 mM Na-Fluorescein (Sigma-Aldrich) was placed in the upper chamber, on top of the cornified layer. The fluorescence intensity in the bottom well was measured after 24 h of incubation with fluorescein (meas. filter 492 nm, ref. filter 570 nm).
Biological Assay
Biotin
Cysteamine
Fluorescein
Fluorescence
Formaldehyde
Keratinocyte
Microscopy, Confocal
Paraffin Embedding
paraform
Permeability
Streptavidin
TERT protein, human
Therapies, Investigational
Vascular Permeability
To calculate the maximum concentration at which there was no cell death a MTS cell viability assay was performed. HUVECs were cultured according to supplier protocols, subcultured, and seeded in a 96 well‐plate at a density of 10 000 cells per well. After allowing for cell adhesion overnight, the cells were treated with serial dilutions of DESs dissolved in fresh HUVEC media. The plate was incubated for 4 hours before the media was aspirated and replaced with fresh media containing 20% MTS Reagent. After 1 hours the absorbance was measured at 490 nm (BioTek Synergy neo2).
To assess vascular permeability, transport across transwell cell culture experiments were used. First, sufficient 24 well‐plate transwell inserts were coated with 0.1% gelatin under sterile conditions and stored at 4 °C overnight. The excess gelatin was removed by inversion of transwell and washed with sterile PBS. HUVECs were cultured according to supplier protocols, subcultured, and seeded on the transwell inserts with 400 µL of 250 000 cells mL−1 media. The outer well was then filled with 600 µL of fresh media and allowed to grow for 48 hours before the experimental progression. After monolayers had formed, the media was removed from the top chamber and replaced with media containing 0.15% IL and 0.9 U mL−1 insulin, this was in preparation for the concentration and insulin to IL ratio to be used in the future in vivo studies. The plate was incubated in appropriate cell culture conditions and 300 µL samples were taken from the plate wells and replaced with fresh media every 10 minutes for 1 hour. The samples were then stored at 4 °C until they were diluted appropriately and quantified using ELISA.
To assess vascular permeability, transport across transwell cell culture experiments were used. First, sufficient 24 well‐plate transwell inserts were coated with 0.1% gelatin under sterile conditions and stored at 4 °C overnight. The excess gelatin was removed by inversion of transwell and washed with sterile PBS. HUVECs were cultured according to supplier protocols, subcultured, and seeded on the transwell inserts with 400 µL of 250 000 cells mL−1 media. The outer well was then filled with 600 µL of fresh media and allowed to grow for 48 hours before the experimental progression. After monolayers had formed, the media was removed from the top chamber and replaced with media containing 0.15% IL and 0.9 U mL−1 insulin, this was in preparation for the concentration and insulin to IL ratio to be used in the future in vivo studies. The plate was incubated in appropriate cell culture conditions and 300 µL samples were taken from the plate wells and replaced with fresh media every 10 minutes for 1 hour. The samples were then stored at 4 °C until they were diluted appropriately and quantified using ELISA.
Biological Assay
Cell Adhesion
Cell Culture Techniques
Cell Death
Cells
Cell Survival
Desmosine
Disease Progression
Enzyme-Linked Immunosorbent Assay
Gelatins
Insulin
Inversion, Chromosome
Sterility, Reproductive
Technique, Dilution
Vascular Permeability
The validated biomechanical model for calf muscle microvascular perfusion was reported previously.15 Briefly, the ANSYS SpaceClaim application within the ANSYS Workbench (Ansys, Inc, v. 18.2) was used for creating 3‐dimensional volumes of leg segments with a thickness of 10 mm to maintain continuity and clinical relevance (same thickness as CE‐MRI scans). The 2‐compartment Tofts‐Kermode model17 of plasma and tissue was taken as the basis of our computational microvascular model, which was used to describe the CE‐MR signal enhancement by GBCA as a reaction term in the tissue regions. The convection‐diffusion and reaction models were adopted to simulate 3‐dimensional convection‐enhanced delivery in the tissue regions.18 The skeletal muscle tissue was modeled as a porous media using the user‐defined model of elastic solid tissue with a porosity of 20% to 80% (Figure ).
The primary microvascular model parameters include the transfer rate constant (kt),15 , 19 , 20 a measure of vascular leakiness; the interstitial permeability to fluid flow (Ktissue)15 , 19 , 20 which reflects the permeability of the microvasculature that is modeled as a porous medium20 ; porosity (φ),15 , 20 , 21 a measure of the fraction of the extracellular space or tissue porosity; the outflow filtration coefficient (OFC)15 , 20 ; and the microvascular pressure (p υ 15 , 20 , 22 The OFC is the product of the vessel permeability and the microvascular surface area per unit volume.15 , 20 Further details of the computational microvascular model were reported previously.15 Parameters of the finite element model were calculated using the programming method implemented in the ANSYS software (v. 18.2). The developed full porous model is a generalization of the Navier–Stokes equations and Darcy law commonly used for flows in porous regions. Blood perfusion velocity and pressure were determined and described for each muscle domain. Blood perfusion was assumed to be transient, incompressible flow. On the boundaries, mass sources were applied to introduce additional fluid into the simulation. The arterial input function corresponding to the signal intensity of the contrast agent in the blood plasma was obtained from CE‐MRI scans1 and applied as a boundary condition for the suggested biomechanical model. Furthermore, a relative static pressure of zero was specified over the outlet boundary, and the flow velocity at the wall boundary was also set to zero. Full information about model parameters is available in our previous publication.15 The grid test and transient time step sensitivity tests were performed to check the quality of the model and for solution convergence. Mean square and mean absolute percentage errors were calculated to estimate the best agreement between simulated and CE‐MRI signal intensities. Intrarater and interrater (for 3 patients with PAD and 5 controls) reproducibility for simulated mean signal intensity over all time steps was excellent for all muscle groups, as reported previously.15
The primary microvascular model parameters include the transfer rate constant (kt),
Arteries
BLOOD
Blood Vessel
Contrast Media
Convection
Diffusion
Elastic Tissue
Extracellular Space
Filtration
Generalization, Psychological
Hypersensitivity
Interstitial Fluid
MRI Scans
Muscle Tissue
Obstetric Delivery
Patients
Perfusion
Permeability
Permeability, Microvascular
Plasma
Pressure
Skeletal Muscles
Step Test
Tissues
Transients
Vascular Permeability
Top products related to «Vascular Permeability»
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The In Vitro Vascular Permeability Assay Kit is a laboratory tool designed to measure the permeability of endothelial cell monolayers in vitro. The kit provides the necessary components to perform quantitative analysis of the barrier function of endothelial cells.
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Evans blue dye is a lab equipment product used as a dye for various research applications. It is a blue azo dye with the chemical formula C₃₄H₂₄N₆Na₄O₁₄S₄. The core function of Evans blue dye is to act as a marker or tracer in biological studies and experiments.
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Evans blue is a dye used as a laboratory reagent. It is a blue-colored dye that binds to albumin in the blood, allowing for the measurement and visualization of blood volume and albumin distribution. The dye has a strong blue color and is soluble in water.
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Formamide is a colorless, odorless, and hygroscopic liquid. It is a common laboratory solvent used in various chemical and biological applications. Formamide has a high boiling point and is miscible with water and many organic solvents.
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FITC-dextran is a fluorescent labeled dextran compound. It is a water-soluble carbohydrate polymer that is covalently linked to the fluorescent dye fluorescein isothiocyanate (FITC). FITC-dextran is commonly used as a tracer or marker in various biological applications.
Sourced in United States, Germany, Switzerland
Texas Red dextran is a fluorescent dye conjugated to a dextran polymer. It is used as a molecular tracer and fluorescent label for various biological applications.
Sourced in United States
The Vascular Permeability Assay Kit is a laboratory instrument used to measure the permeability of vascular endothelial cells. It provides a quantitative assessment of endothelial barrier function.
Sourced in Germany
The in vitro vascular permeability assay is a lab equipment product used to measure the permeability of endothelial cell monolayers. It provides a quantitative assessment of the barrier function of the endothelium.
Sourced in United States, Sao Tome and Principe, United Kingdom
EB dye is a fluorescent compound used in molecular biology and biochemistry applications. It binds to nucleic acids, such as DNA and RNA, and emits an orange-red fluorescence when exposed to ultraviolet or blue light. The core function of EB dye is to enable the visualization and detection of nucleic acids in various lab techniques, such as gel electrophoresis and fluorescence microscopy.
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N,N-dimethylformamide is a clear, colorless liquid organic compound with the chemical formula (CH3)2NC(O)H. It is a common laboratory solvent used in various chemical reactions and processes.
More about "Vascular Permeability"
Vascular permeability is a crucial physiological process that governs the movement of substances, such as fluids, macromolecules, and cells, across the endothelial barrier of blood vessels.
This exchange of nutrients, gases, and waste products between the bloodstream and surrounding tissues is essential for maintaining homeostasis and supporting various biological functions.
Alterations in vascular permeability can contribute to the development of various pathological conditions, including inflammation, edema, and tumor angiogenesis.
Understanding and optimizing the measurement of vascular permeability is crucial for research in fields like cardiovascular biology, oncology, and drug delivery.
Researchers often utilize techniques like the In Vitro Vascular Permeability Assay Kit, Evans blue dye, FITC-dextran, and Texas Red dextran to assess and quantify vascular permeability.
These methods can provide valuable insights into the underlying mechanisms and help identify potential therapeutic targets.
The Evans blue dye, for instance, is a widely used marker for measuring vascular permeability, as it binds to serum albumin and can be detected spectrophotometrically.
Formamide (N,N-dimethylformamide) is commonly used to extract the dye from tissues for quantification.
PubCompare.ai's AI-driven protocol comparison tool can streamline your vascular permeability research by enabling you to easily locate, evaluate, and compare protocols from literature, pre-prints, and patents.
This ensures your studies are grounded in the best available evidence and leverages AI-powered analysis to identify the most accurate and reproducible methods, helping you elevate the quality of your vascular permeability research.
This exchange of nutrients, gases, and waste products between the bloodstream and surrounding tissues is essential for maintaining homeostasis and supporting various biological functions.
Alterations in vascular permeability can contribute to the development of various pathological conditions, including inflammation, edema, and tumor angiogenesis.
Understanding and optimizing the measurement of vascular permeability is crucial for research in fields like cardiovascular biology, oncology, and drug delivery.
Researchers often utilize techniques like the In Vitro Vascular Permeability Assay Kit, Evans blue dye, FITC-dextran, and Texas Red dextran to assess and quantify vascular permeability.
These methods can provide valuable insights into the underlying mechanisms and help identify potential therapeutic targets.
The Evans blue dye, for instance, is a widely used marker for measuring vascular permeability, as it binds to serum albumin and can be detected spectrophotometrically.
Formamide (N,N-dimethylformamide) is commonly used to extract the dye from tissues for quantification.
PubCompare.ai's AI-driven protocol comparison tool can streamline your vascular permeability research by enabling you to easily locate, evaluate, and compare protocols from literature, pre-prints, and patents.
This ensures your studies are grounded in the best available evidence and leverages AI-powered analysis to identify the most accurate and reproducible methods, helping you elevate the quality of your vascular permeability research.