ivis kinetic, by PerkinElmer - Product details

1

Quantifying Tumor Proliferation and Dissemination

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Proliferation and thoracic dissemination of 4T1 cells were quantified by in vivo bioluminescence imaging (IVIS Kinetic, PerkinElmer, Villebon-sur-Yvette, France) on days 7 and 14 following implantation. Five minutes before imaging, vigil mice received an intraperitoneal injection of 150 µg/g of d-luciferin. They were then anesthetized (isoflurane 4% for induction and 1.5% thereafter) and placed in the BLI system (IVIS Kinetic, PerkinElmer, Villebon-sur-Yvette, France), in supine position.
Signal quantifications were carried out by drawing regions of interest on the primary tumor and thorax areas (LivingImage software; PerkinElmer, Villebon-sur-Yvette, France). Results were expressed as photons per second (ph/s).

Minoves M., Kotzki S., Hazane-Puch F., Lemarié E., Bouyon S., Vollaire J., Gonthier B., Pépin J.L., Josserand V., Briançon-Marjollet A, & Godin-Ribuot D. (2022). Chronic intermittent hypoxia, a hallmark of obstructive sleep apnea, promotes 4T1 breast cancer development through endothelin-1 receptors. Scientific Reports, 12, 12916.

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2

Lentivirus-Mediated Lymphoma Induction in Rag1-/- Mice

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For lentivirus production, 293T cells were co-transfected with the viral backbone vector, PMDG and gag-POL packaging vectors donated from Professor Ben-Neriyha's lab using Mirus-TransIT-293 transfection reagent according to standard protocol. EL4 cells were transduced with 5 ml lentivirus in the presence of (6 μg/ml) polybrene. Two days post-infection, puromycin (2.5 μg/ml) was added to the cells for another 1 day at 37°C for the selection of cells containing Slfn2 shRNA or scrambled shRNA. After selection, the cells were infected with the lentivirus containing luciferase donated from Dr. Granot's lab. A total of 6 × 10⁵ infected cells were injected IP into Rag1^−/− mice. The mice were monitored for lymphoma development by the IVIS Kinetic (Caliper Life Sciences) Bioluminescent Fast Imaging system at 13 and 20 days post-injection. Before imaging, the mice were injected with luciferin at 3 mg per mouse (Megapharm, catalog number: 82250).

Goldshtein A., Zerbib S.M., Omar I., Cohen-Daniel L., Popkin D, & Berger M. (2016). Loss of T-cell quiescence by targeting Slfn2 prevents the development and progression of T-ALL. Oncotarget, 7(30), 46835-46847.

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3

In Vivo 2-DG Tumor Imaging

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WT or 5-FU-R cell-bearing nude mice were injected with 10 nmol IRDye 800CW 2-DG Optical Probe (Li-Cor Biosciences) via the tail vein, and then imaged using an IVIS Kinetic (Caliper Life Sciences, USA) small animal imaging system with a cooled Hamamatsu ORCA-R2 camera (Hamamatsu, Japan) 24 h later. Probe signals were displayed as pseudo-colored bioluminescent images and merged with grey-scale white light images of the mice. Circular ROIs were drawn over the areas and quantified, and the results are reported as total radiant efficiency.

Dong S., Liang S., Cheng Z., Zhang X., Luo L., Li L., Zhang W., Li S., Xu Q., Zhong M., Zhu J., Zhang G, & Hu S. (2022). ROS/PI3K/Akt and Wnt/β-catenin signalings activate HIF-1α-induced metabolic reprogramming to impart 5-fluorouracil resistance in colorectal cancer. Journal of Experimental & Clinical Cancer Research : CR, 41, 15.

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4

Xenograft Tumor Monitoring in Nude Mice

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2 × 10⁶ A549 cells or 2.5 × 10⁵ H460 cells expressing luciferase were injected subcutaneously into the right flank of 6-week old female athymic nude mice (Duke Breeding Core, Durham, NC, USA). Tumors were measured with calipers twice a week, and tumor volume was calculated as 0.5 × width² × length. Mice were injected with 100uL of 15mg/mL luciferin and scanned weekly using an IVIS Kinetic (Caliper Life Sciences, Waltham, MA, USA) to assess bioluminescence. Tumors and any metastatic lesions were harvested, and necropsy was performed when tumors reached 2000 mm³ or at end of study. n=12 mice for each cell line for both A549 and H460 experiments were used for a power of 80%. All mice were 6-week old female athymic nude mice so no randomization was performed. Study complies with ethical regulations and approved by the Duke Institutional Animal Care and Use Committee (IACUC). Blinding was accomplished by assigning each mouse a unique identifier without annotation of experimental group. Only this identifier was known to investigators during caliper measurements.

Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.

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5

Evaluating TRAIL-Mediated Cancer Cell Death

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5 × 10³ cancer cells (MDA-MB231-Br or A549) labeled with stable expression of FLuc were seeded in 24-well plates in 10% FBS, 1% penicillin/streptomycin, and 0.01% Plasmocin^™ DMEM. The following day, wells were seeded with varying numbers of hiNSC-TRAIL cells, from 0 to 10,000 cells/well. After 1, 3, and 7 days, viability of the cancer cells was assessed by adding 0.225 μg/mL XenoLight D-luciferin (PerkinElmer) and measuring luminescence using a SynergyH1 microplate reader (BioTek) at 1-sec per well measurement (N=4) or IVIS® Kinetic (Caliper Life Sciences) (N=4–8). Viability was determined by dividing the luminescent signal of these wells by their untreated counterparts at each time point. To test viability of hiNSCs after exposure to recombinant human TRAIL (Sigma), we incubated hiNSCs expressing FLuc with 1–500 ng/mL TRAIL for 48 hours before assessing luminescence on SynergyH1 microplate reader as described above.

Mercer-Smith A.R., Jiang W., Bago J.R., Valdivia A., Thang M., Woodell A.S., Montgomery S.A., Sheets K.T., Anders C.K, & Hingtgen S.D. (2021). Cytotoxic Engineered Induced Neural Stem Cells as an Intravenous Therapy for Primary Non-Small Cell Lung Cancer and Triple-Negative Breast Cancer. Molecular cancer therapeutics, 20(11), 2291-2301.

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6

Bioluminescent Imaging for Tumor and Stem Cell Tracking

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To follow tumor volume or hiNSC-mC-FLuc distribution and persistence, serial bioluminescent imaging (BLI) was performed as previously described (3 (link), 17 ). Mice were administered D-luciferin (3 mg per mouse in 200 μL of PBS) via intraperitoneal injection. 15 min following injection, photon emission was measured using Ami HT (Spectral Instruments Imaging) or IVIS® Kinetic (Caliper Life Sciences). Luminescence was quantified through analysis with Aura (Spectral Instruments Imaging).

Mercer-Smith A.R., Jiang W., Bago J.R., Valdivia A., Thang M., Woodell A.S., Montgomery S.A., Sheets K.T., Anders C.K, & Hingtgen S.D. (2021). Cytotoxic Engineered Induced Neural Stem Cells as an Intravenous Therapy for Primary Non-Small Cell Lung Cancer and Triple-Negative Breast Cancer. Molecular cancer therapeutics, 20(11), 2291-2301.

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7

Fluorescence Imaging and Image-Guided Surgery

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Fluorescence imaging and image-guided surgery were performed with a method described previously.^{20 (link)} Mice were subjected to fluorescence imaging using an IVIS Kinetic optical imaging system (Caliper Life Sciences, Alameda, CA, USA) right after the microPET scan at each time point. The parameters used during the imaging was 10s exposure time (f/stop=4) with an ICG filter set. Image-guided surgery was performed following fluorescence imaging at 48 h after I.V. injection.

Wang M., Mao C., Wang H., Ling X., Wu Z., Li Z, & Ming X. (2017). Molecular Imaging of P-glycoprotein in Chemoresistant Tumors using a Dual-Modality PET/Fluorescence Probe. Molecular pharmaceutics, 14(10), 3391-3398.

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8

In Vivo Tracking of Lipid Nanoparticles

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When the tumor volumes of the SMMC-7721-xenotransplanted BALB/c nude mice reached 200 mm³, LP-ERN-DiR or Tf-LP-ERN-DiR were injected into mice tail veins to give an ERN concentration of 2 mg/kg. The tissue distributions of LP-ERN-DiR and Tf-LP-ERN-DiR in the mice at 2 h, 4 h and 6 h after injection were observed using a small-animal in vivo imaging system (IVIS Kinetic, Caliper, Boston, MA, USA). Finally, the mice were euthanized, tissues were collected, and the fluorescence intensities of tissue types were observed and compared.

Yang A., Sun Z., Liu R., Liu X., Zhang Y., Zhou Y., Qiu Y, & Zhang X. (2021). Transferrin-Conjugated Erianin-Loaded Liposomes Suppress the Growth of Liver Cancer by Modulating Oxidative Stress. Frontiers in Oncology, 11, 727605.

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9

Xenograft Tumor Monitoring in Nude Mice

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2 × 10⁶ A549 cells or 2.5 × 10⁵ H460 cells expressing luciferase were injected subcutaneously into the right flank of 6-week old female athymic nude mice (Duke Breeding Core, Durham, NC, USA). Tumors were measured with calipers twice a week, and tumor volume was calculated as 0.5 × width² × length. Mice were injected with 100uL of 15mg/mL luciferin and scanned weekly using an IVIS Kinetic (Caliper Life Sciences, Waltham, MA, USA) to assess bioluminescence. Tumors and any metastatic lesions were harvested, and necropsy was performed when tumors reached 2000 mm³ or at end of study. n=12 mice for each cell line for both A549 and H460 experiments were used for a power of 80%. All mice were 6-week old female athymic nude mice so no randomization was performed. Study complies with ethical regulations and approved by the Duke Institutional Animal Care and Use Committee (IACUC). Blinding was accomplished by assigning each mouse a unique identifier without annotation of experimental group. Only this identifier was known to investigators during caliper measurements.

Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.

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10

Non-invasive Tumor Imaging Using CB Probe

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When the diameter of tumor xenografts reached 7.0±3.0 mm in size, 2 nmol (100 µl) of the CB probe was injected via tail vein following the probe manual. After injection, the anesthetized mice (n = 6) were placed in the heated imaging platform, the NIRF signals were non-invasively monitored using a living imaging system (IVIS Kinetic, Caliper Life Sciences, USA). Image acquisition parameters were as follows: emission filter = Cy5.5, excitation filter = 675 nm, emission spectra = 680 to 720 nm, exposure time = 0.5 s, f/stop = 2, binning = medium, field of view = 12.5 cm². In vivo imaging of tumor bearing mice were performed every 2 h, 36 h post the CB probe injection, and acquired images were analyzed using living image software (version 4.2.0, Caliper Life Sciences, Inc., US). Fluorescence intensity, defined as total radiant efficiency [p/s]/[μW/cm²], was quantified using identical size regions of interest (ROI). The tumor ROIs were placed with the center over the respective brightest fluorescence in the tumor xenografts. For consistency, the same investigator placed all ROIs. The data were shown as mean ± SD. Normal nude mice (n = 3) acted as control group.

Ma W., Ma L., Zhe H., Bao C., Wang N., Yang S., Wang K., Cao F., Cheng Y, & Cheng Y. (2014). Detection of Esophageal Squamous Cell Carcinoma by Cathepsin B Activity in Nude Mice. PLoS ONE, 9(3), e92351.

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Ivis kinetic

Lab products found in correlation

38 protocols using ivis kinetic

Quantifying Tumor Proliferation and Dissemination

Lentivirus-Mediated Lymphoma Induction in Rag1-/- Mice

In Vivo 2-DG Tumor Imaging

Xenograft Tumor Monitoring in Nude Mice

Evaluating TRAIL-Mediated Cancer Cell Death

Bioluminescent Imaging for Tumor and Stem Cell Tracking

Fluorescence Imaging and Image-Guided Surgery

In Vivo Tracking of Lipid Nanoparticles

Xenograft Tumor Monitoring in Nude Mice

Non-invasive Tumor Imaging Using CB Probe

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