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Intravital Microscopy

Intravital Microscopy is a powerful imaging technique that allows researchers to observe and analyze biological processes in living organisms in real-time.
This non-invasive method enables the visualization of cellular and subcellular dynamics within their native microenvironment, providing invaluable insights into physiological and pathological mechanisms.
By combining advanced optics, fluorescence labeling, and sophisticated image analysis, Intravittal Microscopy has become an indispensable tool in the fields of immunology, cancer biology, neuroscience, and beyond, advancing our understanding of complex biological systems.

Most cited protocols related to «Intravital Microscopy»

1
Fly Visual Perception Correlation Analysis

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Publication 2016
Calcium Diptera Genetic Selection Intravital Microscopy Light Nervousness
2
Intravital Imaging of Tumor Vasculature
See Supplementary Information for extended experimental methods. Complex 1 and 2[23 (link)], [PtCl3NH3]PPh4[24 ], Ag2R(COO)2(R = cyclobutane-1,1-dicarboxylic acid)[35 ] and Amino-BODIPY[36 ] were synthesized as previously described. For 53BP1 transgene expression, a fragment corresponding to amino acids 1220-1711 was subcloned into the pLVX lentiviral vector (Clontech)[26 (link)]. A DeltaVision (Applied Precision) modified Olympus BX63 microscopy system with an environmental chamber was used for live-cell microscopy. Intravital microscopy was performed on an Olympus FV1000 multiphoton imaging system and using animals in accordance with guidelines from the Institutional Subcommittee on Research Animal Care. Following previously described procedures[19 (link)], tumors were injected in nu/nu mice (Cox7, MGH) 30 min following dorsal window chamber implantation and imaged 2 weeks later. Mice were injected via tail vein catheter with Angiosense-680 (Perkin-Elmer) to identify vasculature, 150 nmol CP-11, and imaged.
Miller M.A., Askevold B., Yang K.S., Kohler R, & Weissleder R. (2014). Platinum compounds for high-resolution in vivo cancer imaging. ChemMedChem, 9(6), 1131-1135.
Publication 2014
Amino Acids Animals BODIPY Catheters Cells Cloning Vectors Cyclobutanes Dicarboxylic Acids Intravital Microscopy Mice, Laboratory Mice, Nude Microscopy Neoplasms Ovum Implantation Tail TP53BP1 protein, human Transgenes Veins
3
Murine Model of IVC Flow Restriction
Mice were anesthetized by intraperitoneal injection as described previously (Massberg et al., 2002 (link)). A median laparotomy was performed and the IVC was exposed by atraumatic surgery. We positioned a space holder (FloppyR II Guide Wire 0.014 in [0.36 mm]; Guidant Corporation) on the outside of the vessel and we placed a permanent narrowing ligature (8.0 monofil polypropylene filament, Premilene; Braun) exactly below the left renal vein. Subsequently, the wire was removed to avoid complete vessel occlusion. Side branches were not ligated or manipulated. Flow velocity was determined immediately after the flow restriction (Cap-Image 7.1). Because we wanted to rule out endothelial injury as a trigger for venous thrombosis, all mice with bleedings or any injury of the IVC during surgery were excluded from further analysis. There was no difference in the exclusion rate across the different experimental groups. After the procedure, a subset of animals was investigated by intravital microscopy. In the remainder, the median laparotomy was immediately sutured by a 7.0 polypropylene suture (Ethicon). For weight measurement, the vessel was excised just below the renal veins and proximal to the confluence of the common iliac veins. After the restriction procedure the blood flow velocity was reduced by ∼80% (Fig. 1 B). The shear stress was 0.144 dyne/cm2 ± 0.02 SEM before the flow restriction and 0.072 dyne/cm2 ± 0.017 SEM after the procedure in the IVC close to the site of ligation. Sham experiments consisted of preparation of the IVC and placement of the filament under the vessel without ligation.
von Brühl M.L., Stark K., Steinhart A., Chandraratne S., Konrad I., Lorenz M., Khandoga A., Tirniceriu A., Coletti R., Köllnberger M., Byrne R.A., Laitinen I., Walch A., Brill A., Pfeiler S., Manukyan D., Braun S., Lange P., Riegger J., Ware J., Eckart A., Haidari S., Rudelius M., Schulz C., Echtler K., Brinkmann V., Schwaiger M., Preissner K.T., Wagner D.D., Mackman N., Engelmann B, & Massberg S. (2012). Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. The Journal of Experimental Medicine, 209(4), 819-835.
Publication 2012
Animals Blood Flow Velocity Blood Vessel Cytoskeletal Filaments Dental Occlusion Endothelium Hemorrhage Iliac Vein Injections, Intraperitoneal Injuries Intravital Microscopy Laparotomy Ligation Ligature Mice, House Operative Surgical Procedures Polypropylenes Precipitating Factors Sutures TRAF3 protein, human Vein, Renal Venous Thrombosis
4
Intravital Imaging of Mouse Lymphoid Tissues
Spleens or Peyer’s patches of anesthetized mice were imaged. Spleens or Peyer’s patches were surgically exteriorized, immobilized on a microscope stage, and maintained at 37 °C. A Nikon A1 laser scanning confocal microscope with a 20× objective and software NIS-Elements C was used for image acquisition. We used three dichronic mirrors (DM457/514 and DM405/488/561/640), and three bandpass emission filters (482/35, 540/30, 525/50, 595/50 and 700/75). YFP/CFP ratio was obtained by excitation at 458 nm. PE and Alexa-647 were excited at 488 nm and 633 nm, respectively. Images of purified cells in phosphate-buffered saline were also obtained as above. For 2P microscopy, we used a BX61WI/FV1000 upright microscope equipped with a ×25 water-immersion objective lens (XLPLN25XW-MP; Olympus, Tokyo, Japan), which were connected to a Mai Tai DeepSee HP Ti:sapphire Laser (Spectra Physics, Mountain View, CA). The excitation wavelength for CFP was 840 nm. We used an IR-cut filter BA685RIF-3, three dichroic mirrors (DM450, DM505, and DM570), and three emission filters [FF01-425/30 (Semrock) for the second harmonic generation image, BA460-500 (Olympus) for CFP, and BA520-560 (Olympus) for YFP]. Intravital microscopy of mouse calvaria bone tissues was performed using a protocol modified from a previous study; 10–14-week-old mice were anesthetized using isoflurane; the frontoparietal region of the skull bone was exposed, and the internal surfaces of bones adjacent to the bone marrow cavity were observed using multiphoton excitation microscopy. The imaging system was composed of a multiphoton microscope (A1-MP; Nikon) driven by a laser (Chameleon Vision II Ti: Sapphire; Coherent) tuned to 840 nm together with an upright microscope equipped with a 25× water immersion objective (APO, N.A. 1.1; Nikon). We used three dichronic mirrors (DM458, DM506, and DM561) and three bandpass emission filters (417/60, 480/40, and 534/30). Acquired images were analyzed with MetaMorph software (Universal Imaging, West Chester, PA) and Imaris Software (Bitplane AG, Zürich, Switzerland).
Yoshikawa S., Usami T., Kikuta J., Ishii M., Sasano T., Sugiyama K., Furukawa T., Nakasho E., Takayanagi H., Tedder T.F., Karasuyama H., Miyawaki A, & Adachi T. (2016). Intravital imaging of Ca2+ signals in lymphocytes of Ca2+ biosensor transgenic mice: indication of autoimmune diseases before the pathological onset. Scientific Reports, 6, 18738.
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Publication 2016
Apolipoprotein A-I Bone Marrow Bones Bone Tissue Calvaria Cells Chameleons Cranium Dental Caries Immersion Intravital Microscopy Isoflurane Lens, Crystalline Microscopy Microscopy, Confocal Multiphoton Fluorescence Microscopy Mus Operative Surgical Procedures Peyer Patches Phosphates Saline Solution Sapphire Vision
5
Intravital Microscopy of Cremaster Muscle
2 h before cremaster muscle exteriorization, mice received 4 μg PTx i.v. (Sigma-Aldrich) and 500 ng TNF-α intrascrotally (R&D Systems). Mice were anesthetized with an i.p. injection of 125 mg/kg ketamine hydrochloride (Sanofi), 0.025 mg/kg atropine sulfate (Fujisawa), and 12.5 mg/kg xylazine (TranquiVed; Phoenix Scientific) and placed on a heating pad. The cremaster muscle was prepared as previously described (51 (link)). Postcapillary venules with a diameter between 20 and 40 μm were recorded using an intravital microscope (Axioskop, SW 40/0.75 objective; Carl Zeiss, Inc.) through a digital camera (sensicam qe; Cooke Corporation). Blood flow centerline velocity was measured using a dual-photodiode sensor system (CircuSoft Instrumentation).
Zarbock A., Abram C.L., Hundt M., Altman A., Lowell C.A, & Ley K. (2008). PSGL-1 engagement by E-selectin signals through Src kinase Fgr and ITAM adapters DAP12 and FcRγ to induce slow leukocyte rolling. The Journal of Experimental Medicine, 205(10), 2339-2347.
Publication 2008
Blood Flow Velocity Cremaster Muscle Fingers Intravital Microscopy Iodine-125 Ketamine Hydrochloride Mice, House Sulfate, Atropine Tumor Necrosis Factor-alpha Venules Xylazine

Most recents protocols related to «Intravital Microscopy»

1
Zafirlukast Inhibits Thrombus Formation
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Example 3

The effects of zafirlukast on thrombus formation in mice was determined following laser injury of cremaster muscle arterioles, and observed by intravital microscopy. Male C57/BL6 mouse platelets were labelled with DyLight 649-conjugated anti-GPIb antibody (0.2 μg/g body weight) and either vehicle or zafirlukast (ZFL) infused (at a volume required to achieve a circulating concentration of 20 μM). Following laser injury, images were recorded for 5 minutes. FIG. 5 illustrates the maximum fluorescence intensity of each thrombus formed in vehicle treated mice (n=18 thrombi, circles) or ZFL treated mice (n=12 thrombi, squares) and demonstrates that treatment with ZFL results in a reduction in thrombus size. FIG. 6 illustrates the effects of ZFL on bleeding were determined by tail bleeding assay. Vehicle or ZFL (at a volume required to achieve a circulating concentration of 20 μM) were infused into the femoral veins of C57/BL6 mice, 5 minutes prior to tail biopsy. 0.5 cm of tail tip was excised and blood collected in phosphate-buffered saline (PBS), and time to cessation of bleeding was recorded. Treatment with ZFL was associated with no change in bleeding time. Graphs represent mean±SEM, n=10 per treatment, data analyzed by Student's T test, ***p<0.005.

US11872210B2. Thiol isomerases inhibitors and use thereof (2024-01-16). WESTERN NEW ENGLAND UNIVERSITY [US]. Inventors: Daniel Kennedy [US], Jonathan Gibbins [GB], Lisa-Marie Holbrook [GB].
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Patent 2024
Antibodies, Anti-Idiotypic Arterioles Biological Assay Biopsy BLOOD Blood Platelets Body Weight Cardiac Arrest Cremaster Muscle Fluorescence Injuries Intravital Microscopy Males Mus Phosphates Saline Solution Tail Thrombus Times, Bleeding Vein, Femoral zafirlukast
2
Monitoring Corneal Disease Progression in FK Mice
FK mice were examined daily to monitor the disease progression and photographed with the slit-lamp microscope at 1, 3, 5 dpi. Corneal staining with 2% sodium fluorescein (Feiya Technique Co., Ltd., Jiangsu, China) was used to evaluate the epithelial defect. In vivo confocal microscopy (IVCM) was applied to the diagnosis of FK, which further verified that the mouse model of FK was successfully established. For clinical scores’ assessment, mice were examined at the indicated time to visually grade the disease severity on a scale ranging from 0 to 12.15 (link) The corneal pathology scores were assigned by the following three criteria: area of opacity (0–4), density of opacity (0–4), and surface regularity (0–4). The final corneal clinical score of each mouse was calculated as the sum of the three scores described above.
Xu X., Wei Y., Pang J., Wei Z., Wang L., Chen Q., Wang Z., Zhang Y., Chen K., Peng Y., Zhang Z., Liu J., Zhang Y., Jin Z.B, & Liang Q. (2023). Time-Course Transcriptomic Analysis Reveals the Crucial Roles of PANoptosis in Fungal Keratitis. Investigative Ophthalmology & Visual Science, 64(3), 6.
Publication 2023
Cornea Diagnosis Disease Progression Intravital Microscopy Mice, Laboratory Slit Lamp Examination
3
Intravital Microscopy of TNF-α-Induced Inflammation
Mice were treated by intrascrotal injection of 500 ng TNF-α (R&D Systems) 2 hours prior to microscopy, anesthetized, and prepared for intravital microscopy, as described (60 (link)). Movies from cremasteric postcapillary venules ranging from 20 to 40 μm in diameter were recorded using BX51WI microscope (Olympus) with a water immersion objective ×40, 0.80 NA, and an Olympus charge-coupled device camera (CF8/1, Kappa). Blood samples were taken after the experiments, and WBC and neutrophil counts were determined using ProCyte Dx Hematology Analyzer (IDEXX). Rolling velocity and leukocyte adhesion efficiency (number of adherent cells/mm2 divided by the systemic neutrophil count) were calculated on the basis of the recorded movies using Fiji software (61 (link)). Afterward, cremaster muscles were fixed with 4% PFA (AppliChem) and stained using Giemsa (MilliporeSigma). The number of perivascular cells/mm2 was calculated with a Leica DM2500 microscope equipped with a DMC2900 CMOS camera and an HCX PL APO 100×/1.40 Oil Ph3.
Rohwedder I., Wackerbarth L.M., Heinig K., Ballweg A., Altstätter J., Ripphahn M., Nussbaum C., Salvermoser M., Bierschenk S., Straub T., Gunzer M., Schmidt-Supprian M., Kolben T., Schulz C., Ma A., Walzog B., Heinig M, & Sperandio M. (2023). A20 and the noncanonical NF-κB pathway are key regulators of neutrophil recruitment during fetal ontogeny. JCI Insight, 8(4), e155968.
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Publication 2023
BLOOD Chronic multifocal osteomyelitis Cremaster Muscle Intravital Microscopy Leukocytes Medical Devices Microscopy Mus Neutrophil PL 100 Stain, Giemsa Submersion Tumor Necrosis Factor-alpha Venules
4
Intravital Imaging of Leukocyte Recruitment in Pregnant Mice
Pregnant Lyz2GFP mice (E14.5–E17.5) were anesthetized intraperitoneally with 5 mg/mL ketamine and 1 mg/mL xylazine in 10 mL/kg of normal saline. Two hours prior to intravital microscopy, the uterine horn was carefully exteriorized through an abdominal wall incision followed by the intrauterine injection of 100 μL LPS (1 μg/μL in 0.9% NaCl), 50 μL of TNF-α (10 ng/μL in 0.9% NaCl), or NaCl (0.9%). During the intrauterine injection between 2 fetuses, care was taken not to damage the fetuses or their surrounding yolk sacs. The site of injection between 2 fetuses was marked by a small knot using silk braided suture. Thereafter, the uterine horn was returned, and the abdominal wall incision was temporarily closed by a small metallic clamp. After 2 hours, intravital microscopy (La Vision Biotech and Olympus BX51) to analyze leukocyte recruitment was performed as previously described (8 (link)).
Rohwedder I., Wackerbarth L.M., Heinig K., Ballweg A., Altstätter J., Ripphahn M., Nussbaum C., Salvermoser M., Bierschenk S., Straub T., Gunzer M., Schmidt-Supprian M., Kolben T., Schulz C., Ma A., Walzog B., Heinig M, & Sperandio M. (2023). A20 and the noncanonical NF-κB pathway are key regulators of neutrophil recruitment during fetal ontogeny. JCI Insight, 8(4), e155968.
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Publication 2023
Fetus Intravital Microscopy Ketamine Leukocytes Metals Mice, House Normal Saline Silk Sodium Chloride Sutures Tumor Necrosis Factor-alpha Uterine Cornua Vision Wall, Abdominal Xylazine Yolk Sac
5
Intravital Imaging of Nanoparticle Delivery

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Publication 2023
Animal Model Cremaster Muscle Institutional Animal Care and Use Committees Intravital Microscopy Ion, Bicarbonate Ketamine Males Mice, House Saline Solution Tail Tissues Veins Xylazine

Top products related to «Intravital Microscopy»

1
Bx51wi by Olympus
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The BX51WI is an upright microscope designed for upright water immersion imaging. It features a infinity-corrected optical system and a bright LED illumination system. The BX51WI is suitable for a variety of applications requiring high-resolution imaging in an aqueous environment.
2
Rhodamine 6g by Merck Group
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Rhodamine 6G is a fluorescent dye commonly used in various laboratory applications. It is a synthetic organic compound with a distinctive red-orange color. Rhodamine 6G exhibits strong absorption and emission spectra, making it useful for fluorescence-based techniques such as microscopy, flow cytometry, and immunoassays.
3
Lumpfl wir by Olympus
The LUMPFL-WIR is a specialized lab equipment designed for precise optical measurements. It functions as a luminescence microscope, allowing for the observation and analysis of luminescent samples. The device incorporates a wide-field illumination and detection system to capture comprehensive data from the specimens under investigation.
4
Sytox green by Thermo Fisher Scientific
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SYTOX Green is a nucleic acid stain that is membrane-impermeant, allowing it to selectively label dead cells with compromised plasma membranes. It exhibits a strong fluorescent signal upon binding to DNA.
5
Eclipse fn1 by Nikon
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The Eclipse FN1 is a high-performance optical microscope designed for advanced laboratory applications. It features a durable construction and integrated components for reliable performance.
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C57bl 6 mice by Charles River Laboratories
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The C57BL/6 mouse is a widely used inbred mouse strain. It is a common laboratory mouse model utilized for a variety of research applications.
7
Rostock corneal module by Heidelberg Engineering
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The Rostock Corneal Module is a laboratory equipment designed for corneal imaging and analysis. It provides high-resolution imaging of the cornea, enabling detailed examination and evaluation of corneal structures and features.
8
Axiotech vario by Zeiss
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The Axiotech Vario is a versatile microscope system designed for high-performance imaging applications. It features a modular design, allowing users to customize the system to their specific needs. The Axiotech Vario provides high-quality optics and advanced imaging capabilities to support a wide range of scientific and industrial applications.
9
Matlab by MathWorks
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MATLAB is a high-performance programming language and numerical computing environment used for scientific and engineering calculations, data analysis, and visualization. It provides a comprehensive set of tools for solving complex mathematical and computational problems.
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Lsm 510 meta by Zeiss
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The LSM 510 META is a laser scanning confocal microscope designed for high-resolution imaging. It features a multi-track detection system that enables simultaneous acquisition of multiple fluorescent signals.

More about "Intravital Microscopy"

Intravital Microscopy: Unveiling the Secrets of Living Organisms in Real-Time Intravital Microscopy, a powerful imaging technique, has revolutionized the way researchers observe and analyze biological processes in living organisms.
This non-invasive method allows scientists to visualize cellular and subcellular dynamics within their native microenvironment, providing invaluable insights into physiological and pathological mechanisms.
Through the combination of advanced optics, fluorescence labeling, and sophisticated image analysis, Intravital Microscopy has become an indispensable tool in fields such as immunology, cancer biology, neuroscience, and beyond.
By utilizing specialized equipment like the BX51WI microscope, Rhodamine 6G dye, LUMPFL-WIR objectives, and SYTOX Green nucleic acid stain, researchers can capture high-resolution, real-time images and videos of complex biological systems.
The Eclipse FN1 microscope and C57BL/6 mice, for instance, are commonly used in Intravital Microscopy studies to examine immune responses, tumor dynamics, and neurological processes.
The Rostock Corneal Module and Axiotech Vario systems further expand the capabilities of Intravital Microscopy, allowing researchers to visualize the cornea and other delicate structures in living organisms.
Data analysis and processing are crucial aspects of Intravital Microscopy, and tools like MATLAB and the LSM 510 META confocal microscope play a key role in this regard.
By leveraging these advanced technologies, scientists can extract valuable insights from the wealth of visual data, furthering our understanding of complex biological systems.
PubCompare.ai's AI-driven protocol optimization can help researchers unlock the full potential of Intravital Microscopy.
By comparing the best protocols and products from literature, preprints, and patents, our intelligent comparison tools can enhance the accuracy and efficiency of your research, empowering you to make groundbreaking discoveries.