An additional 21 rats were dedicated to short-term experiments to measure pulmonary CDC retention to provide further insights to our initial retention studies (see Online Supp) in which a different methodology was employed. CDCs were stained with a far-red fluorescent, lipophilic dye (DiD, Thermo Scientific) for 30 min at 37°C and washed to remove free-floating dye. Following CDC infusion, the rats were euthanized at increasing time points (from 1 minute to 72 hours), and the lungs collected in cold PBS. The lungs were imaged using the Xenogen IVIS Imaging system (Ex/Em: 647/665nm; Perkin Elmer) and total radiant efficiency was recorded.
Xenogen ivis imaging system
Manufactured by PerkinElmer
Sourced in United States
The Xenogen IVIS Imaging System is a non-invasive in vivo imaging platform designed for small animal research. The core function of the system is to capture and analyze bioluminescent and fluorescent signals emitted from living subjects.
Automatically generated - may contain errors
Lab products found in correlation
D luciferin, by PerkinElmer (5 mentions)
Living image software, by PerkinElmer (3 mentions)
Steady glo luciferase assay, by Promega (2 mentions)
Xcelligence rtca mp instrument, by Agilent Technologies (2 mentions)
Food pellets, by Inotiv (1 mentions)
Tribromoethanol, by Merck Group (1 mentions)
Nsg mice, by Charles River Laboratories (1 mentions)
Vs110 virtual microscopy system, by Olympus (1 mentions)
Poly 1 c, by Merck Group (1 mentions)
Female ncr nu nu mice, by Taconic Biosciences (1 mentions)
Female severe combined immunodeficiency scid mice, by Charles River Laboratories (1 mentions)
Zetasizer, by Malvern Panalytical (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Athymic nude mice, by Jackson ImmunoResearch (1 mentions)
34 protocols using xenogen ivis imaging system
1
CDC Pulmonary Retention Dynamics
2
In Vivo Bioluminescent Imaging of Listeria Infection
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WT and Trpm2−/− mice were i.v. infected with 108 CFU of L. monocytogenes Xen-32 as previously described (22 (link)). Mice were anesthetized with isoflurane 4 h post-infection and bacterial dissemination was tracked using Xenogen IVIS Imaging System (Perkin Elmer Inc.). Photons were measured during 1 min exposure by keeping the animals in the ventral position. Following the procedure, mice were euthanized and livers and spleens were collected. Bacterial burden within the infected organs was also measured by a 30 s exposure on the Xenogen IVIS system. Photon emissions were quantified with Living Image software (Caliper Life Science).
3
In Vivo Tracking of Hydrogel-Delivered siRNA
4
In-vivo Assessment of Anti-Tumor T-Cell Therapy
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In-vivo experiments were performed in accordance with national and international ethical requirements and were approved by the Italian Ministry of Health (N°88/2016-PR). A previously described NSG mouse model (25 (link), 26 (link)) was used to assess the in-vivo anti-tumour effect of αβ- and γδ-T cells. Seven-ten week-old mice were injected intravenously (i.v.) with Daudi-FF.Luc+ cells (2 × 105/mouse). After tumour engraftment, T cells were injected i.v. (5 × 106 or 2.5 × 107 cells/mouse). Tumour growth was monitored by in-vivo bioluminescence using the Xenogen-IVIS Imaging System (PerkinElmer, Waltham, MA—USA), as previously described (25 (link), 34 (link)). Mice received IL2 and IL15 administrations every 3/4 days for the entire duration of the treatment. Mice were euthanised when the veterinarian detected signs of discomfort or graft-vs.-host disease (GvHD), such as weight loss >15%.
5
Assessing Tumor Growth and Vascular Function in Mice
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LLC or B16F0 cells (1 × 106 cells) were injected s.c. into the right flank of Tie2PEKO and control mice. Mice were killed and tumours were collected at 13 or 10 days post tumour inoculation, respectively. For analysis of tumour growth over time, 1 × 106 B16F10 luciferase-positive cells were injected s.c. into the right flank of mice. Mice were anaesthetized with isoflurane three times a week and luciferase activity was measured 10 min following injection of 2 mg luciferin i.p. with a Xenogen IVIS imaging system (Perkin Elmer). Analysis of barrier function of tumour vessels was investigated as described previously49 (link). Briefly, 15 μl of 100 nm diameter rhodamine-labelled fluorescent latex microsphere particles (LumaFluor Corp, Naples, USA) in 200 μl NaCl solution were injected i.v. 24 h before primary tumour removal. Analysis of vessel perfusion was performed by i.v. injection of 60 mg kg−1 body weight Hypoxy Probe (hpi, HP2-1000 kit) dissolved in 150 μl NaCl solution 45 min before tumour removal. Primary tumours were surgically removed under general anaesthesia. Mice with tumours not effectively removed or with subsequent tumour recurrence were removed from analyses. Mice were killed at the indicated time points or when they showed signs of ill health.
6
In Vivo Biodistribution of Nanobody-FITC
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Nb4 was labeled with FITC (Nb4-FITC). The distribution of Nb4-FITC in nude mice was determined. Mice were anesthetized by inhalation of isoflurane. Ten μL Nb4-FITC (1 mg/kg) was injected via the tail vein. Mice in control group were injected with an equal amount of H1N1-FITC. The distribution of Nb4-FITC and H1N1-FITC was observed at 5 min, 15 min, 30 min, 45 min, 1 h, 2 h, and 3 h after the injection under a Xenogen IVIS Imaging System (PerkinElmer, MA, USA).
7
In Vivo and Ex Vivo Bioluminescence Imaging
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Mice were imaged as previously described12 (link) using a Xenogen IVIS imaging system (Perkin Elmer, Santa Clara, CA), 5 minutes after intraperitoneal injection of D-luciferin (126 mg/kg) dissolved in phosphate-buffered saline (PBS). For ex vivo brain bioluminescence imaging, mice were sacrificed by cervical dislocation immediately after being imaged, decapitated and had their brain dissected. Subsequently, brains were sliced into 1.0-mm thick coronal sections and imaged. The BLI data are reported as raw data, as the total number of counts reaching the charge-coupled device detector.
8
Conditional Gene Expression in Transgenic Mice
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Details of the targeting construct, pBS31’-RBGpA TREtight ColA1, are in the Supplementary Information . A DOX-responsive element controlling SRSF6 expression was targeted downstream of the Cola1 locus by Flp/FRT recombinase-mediated site-specific integration in KH2 ES cells, as described50 (link),51 (link). Injection of targeted ES cells into tetraploid blastocysts to produce fully ES-cell-derived transgenic mice was performed by the CSHL Gene Targeting Shared Resource. DOX was administered to adult mice via food pellets (625 mg/kg) (Harlan Teklad). Hair removal was performed either by shaving, plucking, or applying NAIR lotion for 3 min (Church & Dwight Co) immediately before DOX administration. In vivo imaging of skin GFP expression was measured using a Xenogen IVIS imaging system (Perkin Elmer). No experiments were blinded; each group consisted of gender- and age-matched mice (6 to 52 weeks old C57/BL6/mixed background). As the skin phenotype was not anticipated, we did not perform a power calculation to deduce an adequate group size; instead we used a small group size of 8 treated and 8 control animals, which proved adequate for statistical-significance tests. All mouse experiments were approved by the CSHL Animal Care and Use Committee.
9
Orthotopic Liver Tumor Model in Mice
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Hepa1-6 cells stably expressing firefly luciferase (1.0 × 106 in 100 μL of medium) were orthotopically injected into the livers of recipient mice under anesthesia with tribromoethanol (240 mg/kg, Sigma). The growth of tumors was monitored by bioluminescent imaging with the Xenogen IVIS imaging system (Perkin-Elmer, Fremont, CA, USA). Ten days after being implanted with tumor cells, tumor-bearing mice were evaluated by imaging and randomized into control and treatment groups. Mice were imaged once every 6 days during the course of the treatment.
10
CAR T-cell therapy for Raji-ffluc xenograft
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Six- to eight-week-old male NSG mice purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd were injected via tail vein with 1×106 Raji-ffluc cells. Three days later, 5 mg/mL metformin was dissolved in the drinking water and drank abidingly. The control group drank drinking water. At the same time, each mouse was engrafted with 1×106 CD19-CAR T cells intravenously. For bioluminescence imaging, mice were anesthetized and imaged using a Xenogen IVIS Imaging System (Perkin Elmer Life Sciences, Hopkinton, MA, USA). Bioluminescence imaging was performed on days 3, 10, and 18 posttumor injection. Data were disposed by Living Image® Software, Caliper Life Sciences. The animal study was approved by the Animal Experiment Center of the Second Hospital of Shandong University and followed the Regulations on the management of experimental animals.
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