Qtracker 655 vascular label

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Qtracker™ 655 Vascular Labels are fluorescent probes designed for in vivo cell and tissue labeling. They are composed of a quantum dot nanocrystal core conjugated to a targeting peptide that binds to receptors on the surface of cells. The Qtracker™ 655 labels emit light at a wavelength of 655 nanometers when excited by an appropriate light source.

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Lab products found in correlation

Rhodamine dextran, by Thermo Fisher Scientific (1 mentions) Qtracker 655, by Thermo Fisher Scientific (1 mentions) Leica system, by Leica camera (1 mentions) Rhodamine dextran, by Merck Group (1 mentions) Tcs sp8 dls microscope, by Leica camera (1 mentions)

5 protocols using qtracker 655 vascular label

Intravital Imaging of Vascular Dynamics

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As a GRIN lens system, we used custom singlet GRIN needle microendoscope (length ca. 5.07 mm, diameter = 0.60 mm; NEM-060-10-10-850-S-1.0p, GRINTECH Jena, Germany). The GRIN lens was glued into the lens tubing to a final penetration depth of 650 μm when screwed into the fixator plate. CX3CR1:GFP mice were anesthetized and mounted to the microscope as previously described (10 (link)). Qtracker 655 Vascular Label (Thermo Fisher, MA, US) were injected and images were acquired (505 × 505 px, 500 × 500 μm, unidirectional, line average 4, step size 4.5–6.5 μm, ca. 18 steps, stack time 60 sec). Image stacks were loaded into Imaris 9.3.0 software (Bitplane Zürich, Switzerland), median filter (3 × 3 × 3) was applied. Videos were exported (1024 × 1024, 4 fps) and images were angled maximum intensity projections. Individual mice were measured at 940–950 nm on day 2, 3, 4, 5 after osteotomy.

Stefanowski J., Lang A., Rauch A., Aulich L., Köhler M., Fiedler A.F., Buttgereit F., Schmidt-Bleek K., Duda G.N., Gaber T., Niesner R.A, & Hauser A.E. (2019). Spatial Distribution of Macrophages During Callus Formation and Maturation Reveals Close Crosstalk Between Macrophages and Newly Forming Vessels. Frontiers in Immunology, 10, 2588.

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Multicolor In Vivo Immune Cell Imaging

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Antibodies used for imaging were purchased from BD Biosciences (San Jose, CA, USA) or Thermo Fisher Scientific (Waltham, MA, USA). Labelling of platelets and natural killer (NK) cells was achieved using phycoerythrin (PE)-conjugated anti-mouse CD49b (clone HMα2). Neutrophils were labelled with Brilliant Violet 421-conjugated anti-mouse Ly6G (clone 1A8). CD8+ cells were labelled with eFluor 660-conjugated anti-mouse CD8α (clone 53-6.7). Endothelial cells were labelled using PE-conjugated anti-mouse CD31 (clone 390). Typically, 1–3 ug of each labelling antibody was injected intravenously 10 min before imaging to ensure maximal labelling. Vasculature and perfusion were visualized using either fluorescein (FITC)-conjugate albumin or Qtracker 655 Vascular Label (Thermo Fisher Scientific).

Turk M., Naumenko V., Mahoney D.J, & Jenne C.N. (2018). Tracking Cell Recruitment and Behavior within the Tumor Microenvironment Using Advanced Intravital Imaging Approaches. Cells, 7(7), 69.

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Intravital Multiphoton Imaging of Bone Marrow

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Mice were prepared for intravital TPLSM as previously described^{4 (link)}. TPLSM was performed with a Leica system as described above. EGFP⁺ cells of 7-12 week old female LysM-EGFP mice and tdTomato⁺ cells of Catchup^IVM-red mice were excited at 960nm, at which bone tissue additionally emits a second-harmonic generation (SHG) signal at 480nm. Fluorescent cells were detected with specific filters at 525/50nm (EGFP) or 585/50nm (tdTomato) and SHG was detected via a 460/50nm filter.
Blood flow was visualized by injecting 1.5mg/ml Rhodamine Dextran (Sigma-Aldrich, Cat# R9379-100MG) or 1μM Qtracker™ 655 Vascular Labels (Thermo Fisher, Cat# Q21021MP) in a total volume of 100μl PBS i.v.. Fluorescence was excited at 960nm and detected with a 585/40nm (Rhodamine Dextran) or a 650/50nm (Qtracker 655) filter. Imaging was performed as well in resonant as in non-resonant detection mode. Scan speed was adjusted individually for different vessel types from 600 Hz to 12 KHz.
Neutrophils were activated by injecting 100μg/kg body weight hG-CSF (Neupogen®, Amgen GmbH) i.v. in a total volume of 100μl PBS. The raw data were reconstructed and analyzed using Imaris software (Bitplane) and ImageJ.
Further information on software versions used for data collection and processing are listed in the reporting summary document and Supplementary table 4.

Grüneboom A., Hawwari I., Weidner D., Culemann S., Müller S., Henneberg S., Brenzel A., Merz S., Bornemann L., Zec K., Wuelling M., Kling L., Hasenberg M., Voortmann S., Lang S., Baum W., Ohs A., Kraff O., Quick H.H., Jäger M., Landgraeber S., Dudda M., Danuser R., Stein J.V., Rohde M., Gelse K., Garbe A.I., Adamczyk A., Westendorf A.M., Hoffmann D., Christiansen S., Engel D.R., Vortkamp A., Krönke G., Herrmann M., Kamradt T., Schett G., Hasenberg A, & Gunzer M. (2019). A network of trans-cortical capillaries as mainstay for blood circulation in long bones. Nature metabolism, 1(2), 236-250.

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Microangiography for Vascular Leakage Imaging

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Microangiography was performed following a previously published protocol (Arnold et al., 2022 (link); Shin et al., 2019 (link)). Embryos were anaesthetised with Tricane and injected with 1 nl of Qtracker™ 655 Vascular labels (Thermo Fisher Scientific) at 7 dpf in the LFL for valve leakage experiments and imaged on a Leica TCS SP8 DLS microscope at ∼5 min post-injection.

Panara V., Yu H., Peng D., Staxäng K., Hodik M., Filipek-Gorniok B., Kazenwadel J., Skoczylas R., Mason E., Allalou A., Harvey N.L., Haitina T., Hogan B.M, & Koltowska K. (2024). Multiple cis-regulatory elements control prox1a expression in distinct lymphatic vascular beds. Development (Cambridge, England), 151(9), dev202525.

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Microangiography of Facial Lymphatics

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Microangiography was performed as previously (Shin et al., 2016b (link)), except that Qtracker 655 Vascular Labels (Qdot655, ThermoFisher) were injected between the gill and pectoral fin into the facial lymphatic system of anesthetized TgBAC(dab2:egfp)^ncv67Tg embryos. Embryos with successful and specific perfusion of the caudal region of the facial lymphatic vessel were identified using an MZFLIII fluorescence dissection microscope and imaged by confocal microscopy. After imaging, each embryo was given a unique identifier number and transferred into egg water in a single well of a 24-well plate with or without 2x Tricaine. After 30 minutes, the embryos were re-anesthetized and subjected to confocal microscopy.

Shin M., Nozaki T., Idrizi F., Isogai S., Ogasawara K., Ishida K., Yuge S., Roscoe B., Wolfe S.A., Fukuhara S., Mochizuki N., Deguchi T, & Lawson N.D. (2019). Valves are a conserved feature of the zebrafish lymphatic system. Developmental cell, 51(3), 374-386.e5.

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