Live imaging was done weekly by using the Xenogen in vivo imaging system, IVIS-100 (Perkin Elmer, MA, USA) to monitor the orthotopic tumor growth kinetics of the luciferase-labelled ESCC cell lines injected into the mice and to observe for metastasis. The 3D live images were captured by using the Xenogen IVIS Spectrum. Luciferin substrate (Perkin Elmer) at 150 mg/kg was injected into the animals prior to bioluminescence imaging. Animals were euthanized at the end of the study at weeks 3 to 5 to excise the orthotopic tumors. The tumors were dissected and fixed in formalin and embedded in paraffin. Sectioned tissues were stained with hematoxylin and eosin for histological examination by pathologists (Kwok Wah Chan and Alfred K. Lam).
Ivis 100
Manufactured by PerkinElmer
Sourced in United States
The IVIS 100 is an in vivo imaging system designed for real-time visualization and quantification of bioluminescent and fluorescent signals in small animal models. It provides high-sensitivity detection and high-resolution imaging capabilities for a variety of preclinical research applications.
Automatically generated - may contain errors
Lab products found in correlation
D luciferin, by Biosynth (4 mentions)
Living image software, by PerkinElmer (4 mentions)
Ivis lumina, by PerkinElmer (3 mentions)
D luciferin, by GoldBio (3 mentions)
Probumin, by Merck Group (2 mentions)
Forskolin, by Merck Group (2 mentions)
Luciferin, by Promega (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Luciferin substrate, by PerkinElmer (1 mentions)
F 12k nutrient mixture, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Ami imager, by Spectral Instruments Imaging (1 mentions)
D luciferin potassium salt, by PerkinElmer (1 mentions)
Ivis 50, by PerkinElmer (1 mentions)
D luciferin, by PerkinElmer (1 mentions)
D luciferin, by Thermo Fisher Scientific (1 mentions)
Ivis spectrum ct, by PerkinElmer (1 mentions)
30 protocols using ivis 100
1
Longitudinal Live Imaging of Orthotopic ESCC Tumors
2
Luminescent Cell-Based Assay for Enzyme Activity
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Chinese hamster ovary (CHO) cells and HeLa cells were grown in a CO2 incubator at 37°C with 5% CO2 and were cultured in F-12K Nutrient Mixture (GIBCO) and Dulbecco’s Modified Eagle’s Medium (DMEM) (GIBCO) respectively. Both media were supplemented with 10% fetal bovine serum and 100 U/mL penicillin/streptomycin. CHO cells were transfected with pcDNA3.1 plasmids containing either Fluc, CG6178, or FruitFire. Cells were plated into 96-well black plates (Costar 3915) at 10k cells per well. After 24 hours, cells were transfected with 0.075 μg DNA per well using Lipofectamine 2000 (ThermoFisher). Prior to imaging the transfected cells, a master plate containing the substrates titrated from 250 μM to 0.122 μM over the 12 columns was prepared at sufficient volume to supply substrate to the plates for all three enzymes being assayed. At 24 hours post-transfection, cells were imaged by removing the media containing the Lipofectamine, washing each well with 60 μl Hank’s Balanced Salt Solution (HBSS), aspirating the HBSS, then adding 60 μl per well of the appropriate substrate. After a 3 minute hold, the cells were imaged on a Perkin Elmer IVIS 100. Data collection and Regions of Interest (ROIs) were performed with Living Image Software, then exported to Excel for formatting. The formatted data was plotted using GraphPad Prism and analyzed using nonlinear regression models.
3
Raji Lymphoma Model for NK Cell Therapy
4
Quantifying CXCL12 Isoform Effects on cAMP
5
Quantifying CXCL12 Isoform Effects on cAMP
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To measure CXCL12-dependent effects on intracellular cAMP, we plated 2 × 104 231-CXCR4 cells stably expressing the firefly luciferase cAMP reporter (231-CXCR4-cAMP) in black wall 96 well plates one day before assays. We incubated cells for 10 minutes with equal amounts of CXCL12-α, β, γ; unfused GL; or DMEM with 0.2% probumin (Millipore). We then added 5 μM forskolin (Sigma) to increase intracellular cAMP or vehicle control in combination with 150 μg/ml luciferin (Promega) for imaging on an IVIS 100 (Perkin-Elmer, Waltham, MA, USA). Images (large binning, four minute acquisition) were obtained 12 minutes after adding forskolin.
We quantified recruitment of β-arrestin 2 to CXCR4 in response to different isoforms of CXCL12 fused to GL or GL control as described (28 (link)).
We quantified recruitment of β-arrestin 2 to CXCR4 in response to different isoforms of CXCL12 fused to GL or GL control as described (28 (link)).
6
Tamoxifen-Induced Osteoblast Tracking
7
Bioluminescence Imaging of Infected Mice
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Groups of infected mice were subjected to bioluminescence analysis under isofluorane anaesthetic using an IVIS 100 (Perkin Elmer, MRC Biological Imaging Centre) or IVIS Lumina (Perkin Elmer, kind access Lung Immunology Group). During imaging, a reference image was obtained of the animal under low illumination prior to quantification of photons emitted from bioluminescent GAS strains at a binning of 4 over 60 s. Photon emission was determined using the Living image Software package (Version 3.2) and region of interests (ROI) were calculated and displayed as total flux and then compared to bacterial counts in the same organ tissue.
8
In-Vivo Bioluminescence Imaging of Breast Cancer
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IVIS® 100, a commercial BLI system (PerkinElmer, Inc. Waltham, MA) was used to acquire luminescent images from breast cancer-bearing mice in-vivo. Images were collected; (1) at 72 h after cancer cell implantation, (2) just prior to ALA injection, and (3) at 24 h after PDT. Prior to in-vivo imaging, mice were anesthetized with inhaled 1–3% isoflurane. They were then given d -luciferin (150 mg/kg subcutaneously, in Dulbecco's phosphate buffered saline, PBS) and placed on a warm (37°C) stage inside the light-tight camera box, under continuous 1–3% isoflurane anesthesia. Three or four mice were imaged at the same time. Camera settings were: Field of View, B; Exposure time, 20 sec; Binning, medium; F/stop, 1. Digital images were obtained 12–15 min after d -Luciferin injection. Images from multiple days were normalized and analyzed using LivingImage® 4.2 software (Xenogen, Alameda, CA, USA); the emitted light in each region of interest (ROI) was quantified as total photon counts, or radiance (see Fig.1 ).
9
In Vivo Tracking of Stem Cell Engraftment
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FACS sorted and expanded Luciferase+GFP+ hBM-MSCs were injected into the wound beds or non-wounded skin of anesthetized db/db mice or NSG mice (2.5 × 105 cells/mouse) on the day of wounding unless otherwise indicated. At different timepoints post hMSC injection, luciferase activities were detected in vivo. Mice were injected intraperitoneally with d -Luciferin (0.15 g luciferin per 1 g bodyweight) and imaged 10 min after with IVIS-100 or IVIS Spectrum imaging system (Perkin Elmer, Waltham, MA), or with Ami Imager (Spectral Instruments Imaging, Tucson, AZ, USA) based on instrument availability in different animal housing facilities. Images were obtained and analyzed using manufacturers’ software (Perkin Elmer: LivingImage; Spectral Instruments Imaging: Aura). Bioluminescence was quantified using the LivingImage software (Perkin Elmer).
10
Quantifying Transplant Survival in Rats Using IVIS Imaging
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To examine transplant survival in rats over time, and to assess whether IVIS imaging could be used to measure transplant survival more rapidly than stereological quantification, changes in light emission over the first 7 d following transplantation were quantified using the IVIS 100 (PerkinElmer). Initial experiments examined the sensitivity of the IVIS for detecting different numbers of transplanted cells and compared intraperitoneal injection of D-luciferin with intravenous injection following transplantation of 2 × 106 GFP-luc SCs. Intravenous administration via the tail vein resulted in a more rapid increase in light production than intraperitoneal administration (data not shown). The time course of the decrease in light emitted from the transplants over time was compared to the number of transplanted cells at 3 d (n = 4) and 7 d (n = 5). A subset of these rats were quantified by stereology to determine the number of surviving SCs at 3 d (n = 3) and 7 d (n = 4). Spinal cords from two of the rats that underwent IVIS imaging were not available for histologic analysis.
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