The Gr-1+CD11b+ myeloid cells were sorted from spleens of GFP mice, and injected into the tail vein after the window frame was installed in the diabetic mice. After 24 hrs, the mice were injected intravenously with a solution of high molecular weight dextran tetramethylrhodamine (Molecular Probes, Eugene, OR, USA). Immediately after the injection of dextran, the homing of Gr-1+CD11b+ myeloid cells to the site of injury and vasculature was observed under the confocal microscope (Zeiss LSM 510, Munchen, Germany) through the window chamber and photographed.
Tetramethylrhodamine dextran
Manufactured by Thermo Fisher Scientific
Sourced in United States, France, Belgium
Tetramethylrhodamine dextran is a fluorescently-labeled dextran compound. It is used as a tracer molecule in biological and biochemical research applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510, by Zeiss (4 mentions)
Dextran fluorescein, by Thermo Fisher Scientific (3 mentions)
D1817, by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Ms 222, by Merck Group (2 mentions)
Tcs sp8 mp flim, by Leica (2 mentions)
Acridine orange, by Merck Group (2 mentions)
Lucifer yellow ch, by Merck Group (2 mentions)
Alexa fluor 633 hydrazide, by Thermo Fisher Scientific (2 mentions)
X40 0.8 n a, by Olympus (2 mentions)
W water immersion objective, by Olympus (2 mentions)
Tetramethylrhodamine dextran, by BMG Labtech (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Sucrose, by Avantor (1 mentions)
Agarose, by Thermo Fisher Scientific (1 mentions)
Fibronectin, by Merck Group (1 mentions)
Gelatin, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
57 protocols using tetramethylrhodamine dextran
1
In Vivo Tracking of Myeloid Cells
2
Tracking Myeloid Cell Homing in Diabetes
3
In Vitro Axon Guidance Assay
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Agarose (Fischer Scientific, BP1356–100), Bovine Serum Albumin (Sigma, A4503), Fibronectin (Sigma, F1141), dextran fluorescein (Invitrogen, D1820), dextran tetramethylrhodamine (Molecular Probes life technologies, D3312), Gelatin (Sigma, G1890), Methylcellulose (Sigma, M0387–100), Mowiol 40–88 (Fluka, 81386), Penicillin-Streptomycin (Sigma, P4458), Sucrose (VWR, 27480.294). Danilchick’s medium 1 × (DFA): NaCl (53 mM), NA2CO3 (5 mM), K Gluconate (4.5 mM), Na Gluconate (32 mM), MgSO4–7H20 (1 mM), CaCl2 (1 mM), BSA 0.1%. MEMFA: MOPS (1 mM), EGTA (2 mM), MgSO4 (1 mM), Formaldehyde (3.7%). Normal Amphibian Medium (NAM): NaCl (110 mM), KCl, (2 mM), Ca(CO3)2 (1 mM), MgSO4 (1 mM), EDTA (0.1 mM), NaHCO3 (1 mM), Sodium Phosphate (2 mM).NTMT: NaCl (0.1 M), TrisHCl pH9.5 (0.1 M), MgCl2 (50 mM), Tween 0.1%. Phosphate Buffer Saline 1 × : NaCl (137 mM), KCl (2.7 mM), Na2HPO4 (10 mM), KH2PO4 (1.8 mM), CaCl2–2H2O (1 mM), MgCl2–6H2O (0.5 mM). Human Sdf1 (Calbiochem, 572300; used at 0.5 μg/mL in solution). Mouse Semaphorin-3A-Fc (R&D Systems, 5926-S3; used at 15, 30 or 60 ng/mL coated or added in solution), mouse Semaphorin-3F-Fc (R&D Systems, 3237-S3; used at 120, 240 or 480 ng/mL coated).
4
Visualizing Intestinal Vasculature and Cells
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We rendered intestinal tissue transparent using Focus Clear (CelExplorer Labs Co, Hsinchu, Taiwan), according to the manufacturer’s instructions. Briefly, anesthetized mice were injected intravenously with Hoechst 33342 (Thermo Fisher Scientific, Waltham, MA) and dextran-tetramethylrhodamine (Thermo Fisher Scientific). After 30 minutes, mice were killed and the small intestinal tissue was collected, fixed, and incubated in Focus Clear. Samples were mounted on stage with Mount Clear (CelExplorer Labs Co). Z-stack images were taken at intervals of 0.425 μm using an A1RMP microscope (Nikon, Tokyo, Japan). Three planes were chosen at intervals of 131 slices, and GFP+ cells were detected automatically according to the algorithm of Imaris software (Bitplane, Zurich, Switzerland). The number of cells in contact with blood vessels was counted manually.
5
Visualizing Cerebral Vasculature in Mice
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deeply anesthetized mice were injected with dextran-tetramethylrhodamine (Thermo Fisher Scientific, cat no. D1817) 2 mg/20 g body weight to the left ventricle and it was allowed to circulate in the blood stream for 5–10 min. After perfusion, brain was dissected, fixed and treated as described for immunohistochemistry.
6
Tracing Vascular Permeability in CNS
7
Cellular Uptake of Liposomal QS-21
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RAW 264.7 cells were cultured and kept at 37°C in a humidified atmosphere containing 5% CO2. At 70% confluency, cells were incubated for 16 hours with Lucifer yellow (1 mg ml−1; Thermo Fisher Scientific, catalog no. L1177), Dextran, Alexa Fluor 647; 10,000 MW (50 μg ml−1; Thermo Fisher Scientific, catalog no. D22914), and Dextran, Tetramethylrhodamine, 40,000 MW (50 μg ml−1; Thermo Fisher Scientific, catalog no. D1842) and then washed with PBS before incubating the cells for 4 hours with QS-21 (10 μg ml−1) equivalent of either CPQ liposomes (QS-21 containing liposomes) before or after lyophilization. Liposomes lacking QS-21 was used at the same total lipid concentration in the same conditions as a control. The cells were lastly washed and incubated with Hoechst 33258, Pentahydrate (bis-Benzimide) (2.5 μg ml−1; Thermo Fisher Scientific, catalog no. H3569) before imaging using confocal microscopy at 63× magnification power.
8
Zebrafish Embryo Heparinase I Injection
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Heparinase I from Flavobacterium heparinum (Merck) was dissolved in PBS to 1 U/μl and stored as frozen aliquots. For microinjection into the BV, MS-222 (Merck) anesthetized 48hpf zebrafish embryos were laterally mounted in 1% low gelling agarose (Merck). Reaction mix containing 0.1 U/μl heparinase I and 70,000 MW Dextran-Tetramethylrhodamine (ThermoFisher Scientific) was injected into the fourth ventricle of immobilized embryo. Injected embryos were freed from agarose and allowed to recover in glass bottomed dishes prior to imaging.
For surfen treatment, 24 hpf embryos were incubated in 3 µM surfen hydrate (Merck) in 1% DMSO in accordance with Naini et al., 2018 (link) for 24 hr and measured at 48 hpf.
For surfen treatment, 24 hpf embryos were incubated in 3 µM surfen hydrate (Merck) in 1% DMSO in accordance with Naini et al., 2018 (link) for 24 hr and measured at 48 hpf.
9
Tracing Vascular Permeability in CNS
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Dextran-Tetramethylrhodamine tracer (Dextran-Tetramethylrhodamine, 10-kDa, D1817, ThermoFisher) was injected into mice through tail vein injection, and allowed to circulate for 6 hours. Brain and spinal cord were then dissected immediately and fixed by immersion fixation overnight in 4% PFA to immobilize the tracers at the end of the experiment. The Dextran-Tetramethylrhodamine tracer leakage was assessed within frozen sections.
10
Inhibitor-Based Macropinocytosis Assay
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Inhibitors: PI3K inhibitors: GDC-0941 (HY-50094, used at 5 µM) and ZSTK474 (HY-50847, used at 5 µM), AXL inhibitors: R428 (HY-15150, used at 5 µM) and LDC1267 (HY-12494, used at 5 µM), AKT inhibitor: MK-2206 (HY-10358, used at 0.5 µM) all from MedChemExpress; macropinocytosis inhibitor 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) from Sigma-Aldrich (A3085, used at 25 µM). Other: vitamin K 1 (3804.1, Carl Roth GmbH), puromycin (Toku-E, P001, used at 1 µg/mL), Geneticin® Selective Antibiotic (G418, 11811031, used at 1 mg/mL), Dextran, Tetramethylrhodamine, 70,000 MW (D1818, used at 1 mg/mL), Dextran, Texas Red™, 70,000 MW (D1864, used at 0.25 mg/mL), Dextran, Oregon Green™ 488, 70,000 MW (D7173, used at 0.25 mg/mL) all from Thermo Fisher Scientific; fibronectin (F1056, used at 20 μg/mL), Bovine Serum Albumin (A1470), DAPI (D9542), phalloidin-Atto 390 (50556) all from Sigma-Aldrich.
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