Living image

Manufactured by PerkinElmer

Sourced in United States

Living Image is a bioluminescence and fluorescence imaging system designed for small animal research. The system captures and analyzes images of light-emitting or light-reflecting biological samples, such as cells or tissues, to visualize and quantify biological processes in living subjects.

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133 protocols using living image

Monitoring Metastatic Burden with Bioluminescence

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Metastatic burden in live animals was monitored with bioluminescence imaging (melanomas were tagged with stable expression of luciferase). Five minutes prior to imaging, mice were injected intraperitoneally with 100 μl of PBS w/ D-luciferin monopotassium salt (40 mg/ml, Biosynth). Mice were anesthetized with isoflurane just prior to imaging. All mice were imaged with an IVIS Imaging System 200 series (Caliper Life Sciences) with Living Image software. The bioluminescence signal (total photon flux) was quantified with the “region of interest” tool in Living Image (Perkin Elmer) software.

Gill J.G., Leef S.N., Ramesh V., Martin-Sandoval M.S., Rao A.D., West L., Muh S., Gu W., Zhao Z., Hosler G.A., Vandergriff T.W., Durham A.B., Mathews T.P, & Aurora A.B. (2022). A short isoform of spermatogenic enzyme GAPDHS functions as a metabolic switch and limits metastasis in melanoma. Cancer research, 82(7), 1251-1266.

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In vivo NIRF Imaging of CRANAD-29

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The reagents used for the synthesis were purchased from Aldrich and used without further purification. CRANAD-29 was synthesized according to our previously reported procedures [52 (link)]. All animal experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at Massachusetts General Hospital and carried out in accordance with the approved guidelines. In vivo NIRF imaging was performed using the IVIS® Spectrum animal imaging system (Caliper Life Sciences, Perkin Elmer, Hopkinton, MA), and data analysis was conducted using Living Image® 4.2.1 software. Mice were anesthetized with isoflurane balanced with oxygen during image acquisition (less than 5 minutes for each imaging session).

Yang J., Yang J, & Ran C. (2018). Spectral Unmixing Imaging for Differentiating Brown Adipose Tissue Mass and Its Activation. Contrast Media & Molecular Imaging, 2018, 6134186.

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Murine Xenograft Tumor Model

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Female nude mice were obtained from Jackson Laboratory. Experimental protocols were approved by the Institutional Animal Care and Use Committee at Wake Forest health Science. Female NU/NU Nude Mice from Charles River Laboratories were used for animal experiments. Mice were housed in a 12h light/12 h dark cycle with temperature-controlled room. The animal rooms are provided with 100% fresh, HEPA filtered air at 10-15 air changes per hour. Room temperatures are controlled by reheat units within each room, and are maintained within the range of 70°F ± 2° F. The humidity levels are controlled globally, and it is maintained between 30-70%. The mice were fed with a standard chow (Prolab Isopro RMH 3000, 5P00, LabDiet) and water ad libitum. Approximately 1 million cells were subcutaneously injected into the mammary fat pad of 7- to 9-week-old mice. Tumor length (L) and width (W) were measured by caliper, and tumor size was calculated using the formula, 1/2* LW^{2 (link)}. The maximum tumor size is 2cm of the length and maximal tumour size/burden was not exceeded in this study. Tumor growth were also monitored by bioluminescence imaging using the IVIS lumina III in vivo imaging system (Perkin Elmer). Living Image (Caliper Life Science) version 4.7.3 was used to analyze the bioluminescence level. Tumor wet weight were measured at the endpoint.

Zhao D., Wu K., Sharma S., Xing F., Wu S.Y., Tyagi A., Deshpande R., Singh R., Wabitsch M., Mo Y.Y, & Watabe K. (2022). Exosomal miR-1304-3p promotes breast cancer progression in African Americans by activating cancer-associated adipocytes. Nature Communications, 13, 7734.

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Bioluminescence Imaging of Glioma Cells

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shIDH1-expressing GIC-20, GIC-387, and SUDHL4 and IDH1-overexpressing
NSC-2201 cells were lentivirally transduced with a cDNA encoding firefly
luciferase. Upon cell implantation, mice were injected with luciferin potassium
salt, anesthetized, and tumor growth was monitored by bioluminescence imaging
(IVIS Spectrum, PerkinElmer). Bioluminescence was analyzed using Living Image
(Caliper Life Sciences) software, and bioluminescence was quantified relative to
non-tumor bearing mice.

Calvert A.E., Chalastanis A., Wu Y., Hurley L.A., Kouri F.M., Bi Y., Kachman M., May J.L., Bartom E., Hua Y., Mishra R.K., Schiltz G.E., Dubrovskyi O., Mazar A.P., Peter M.E., Zheng H., James C.D., Burant C.F., Chandel N.S., Davuluri R.V., Horbinski C, & Stegh A.H. (2017). Cancer-associated IDH1 promotes growth and resistance to targeted therapies in the absence of mutation. Cell reports, 19(9), 1858-1873.

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In Vivo Bioluminescent Imaging of E. coli

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At specified times following infection with E. coli Xen 14, in vivo bioluminescent imaging was performed using an in vivo imaging system (IVIS Spectrum, PerkinElmer). CD-1 mice were used for these studies because of the increased sensitivity of imaging due to their white fur. Mice were anesthetized with isoflurane and imaged for 1 to 60 seconds. Bioluminescent flux, measured in photons per second emitted from specific regions, was quantified using computer software (LivingImage, Caliper Life Sciences, Hopkinton, MA); luciferase activity is expressed as photons per second.

Prestia K., Bandyopadhyay S., Slate A., Francis R.O., Francis K.P., Spitalnik S.L., Fidock D.A., Brittenham G.M, & Hod E.A. (2014). Transfusion of stored blood impairs host defenses against Gram-negative pathogens in mice. Transfusion, 54(11), 2842-2851.

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In Vivo Fluorescence Imaging of ICG-Glu-Glu-AE105 in Mice

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Mice (n = 3) were injected intravenously with 10 nmol ICG-Glu-Glu-AE105 and subjected to dynamic imaging at 1, 2, 4, 8, 12, 24, 48, and 72 h. Before scanning, the mice were anaesthetized with 2% isoflurane and positioned in the prone position. For imaging IVIS Lumina XR and acquisition software Living Image (Caliper life Sciences, Hopkinton, CA, USA) were used. The excitation filter was set to 710 nm and the emission filter was set in the ICG. Acquisition was recorded in auto-setting to achieve the best acquisition possible.

Juhl K., Christensen A., Persson M., Ploug M, & Kjaer A. (2016). Peptide-Based Optical uPAR Imaging for Surgery: In Vivo Testing of ICG-Glu-Glu-AE105. PLoS ONE, 11(2), e0147428.

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In Vivo Imaging of ICG-Liposomes

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Mice were shaved under isoflurane (1-2%, v/v) anesthesia and underwent baseline scan using an IVIS Kinetic in vivo imaging system (Caliper Life Sciences, Hopkinton, MA, USA). Two hundred µL of ICG-liposomes or the free compound (8 mg/kg) in sucrose buffer (2 mM phosphate buffer pH 7.0, with 9.3 % sucrose) were injected into the tail vein and the mice were repetitively scanned. Image analysis was performed using Living Image (Caliper Life Sciences, Hopkinton, MA, USA).

Portnoy E., Vakruk N., Bishara A., Shmuel M., Magdassi S., Golenser J, & Eyal S. (2016). Indocyanine Green Liposomes for Diagnosis and Therapeutic Monitoring of Cerebral Malaria. Theranostics, 6(2), 167-176.

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Bioluminescence and Fluorescence Imaging of Tumor Response to PTT

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Animals were scanned for bioluminescence using the IVIS Lumina XR (Caliper life Sciences, Hopkinton, CA, USA) a day before PTT (day -1), on PTT day (day 0) and afterwards every other day until endpoints were reached. For this, mice were injected with luciferin intraperitoneally (5 μl/mg of body weight, at a concentration of 150 mg/mL) ten minutes before the scan. Animals were then anesthetized and placed in the prone position. The bioluminescent signal was quantified within the acquisition software Living Image (Caliper Life Sciences, Hopkinton, CA, USA) by drawing ROIs on the tumors and obtaining the photon flux (photons/s/cm²).
The fluorescence scans were performed to detect ICG signal before and after PTT using the Fluobeam^®800 NIR-camera as described for the accumulation study.

Simón M., Jørgensen J.T., Juhl K, & Kjaer A. (2021). The use of a uPAR-targeted probe for photothermal cancer therapy prolongs survival in a xenograft mouse model of glioblastoma. Oncotarget, 12(14), 1366-1376.

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In Vivo Leukemia Bioluminescence Imaging

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Bioimaging of leukemia burden in vivo was performed by Xenogen IVIS Spectrum system (Caliper Life Science). Before performing, Luciferin (in vivo grade, VWR) was prepare in DPBS, and 150 mg/kg Luciferin was injected by i.p., after 10 min, luminescence signal can be detected for different time points. The signal data was analyzed by the Living Image (Caliper Life Science) software.

Sheng Y., Yu C., Liu Y., Hu C., Ma R., Lu X., Ji P., Chen J., Mizukawa B., Huang Y., Licht J.D, & Qian Z. (2020). FOXM1 regulates leukemia stem cell quiescence and survival in MLL-rearranged AML. Nature Communications, 11, 928.

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Quantifying Tissue Biodistribution of n-hfPNA/Alexa546 in PDX Tumors

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Mice bearing F2 PDX tumors were dosed with n-hfPNA/Alexa546 (10 mg/kg) by tail vein injection. Mice were euthanized at 1, 4, 8 and 24 h post injection to collect lung, liver, kidney, spleen, and PDX tumor tissue samples. PBS was used as a control. The fluorescence of n-hfPNA/Alexa546 in tissue samples was quantified by ex vivo imaging with IVIS. Fluorescence density maps were created for PNAs in each tissue sample using Living Image (Caliper Life Sciences, Hopkinson, MA), and the maps were represented as a color gradient centered at maximal fluorescence intensity.
PDX tumor tissue samples were further analyzed by fluorescence microscopy. Tissue samples were fixed in 4% paraformaldehyde solution containing 10% sucrose at 4°C. The fixed tissue samples were embedded in OCT on dry ice, sectioned at 10 μm thickness, and dried overnight in the dark. The tissue samples were washed with PBS, stained with Hoechst 33342 (or DAPI), and put in an aqueous permanent mounting medium. PDX tumor tissue samples were also treated with EpCAM or CD31 antibodies and NL637-labelled IgG secondary antibodies to visualize cancer cells and blood vessels for fluorescence imaging with a laser scanning confocal microscope (Olympus FV1000, Tokyo, Japan).

Reichel D., Curtis L.T., Ehlman E., Evers B.M., Rychahou P., Frieboes H.B, & Bae Y. (2017). Development of Halofluorochromic Polymer Nanoassemblies for the Potential Detection of Liver Metastatic Colorectal Cancer Tumors Using Experimental and Computational Approaches. Pharmaceutical research, 34(11), 2385-2402.

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