M2-Mφ or MSC-EVs were labeled with sulfo-Cy7-NHS ester according to the manufacturer's instructions (New Research Biosciences Co., Ltd., Xi'an, China). The unbound dyes were removed using an Exosome Spin Column (MW3000, Invirogen). Cy7-labeled M2-EVs or MSC-EVs (Cy7-EVs, 80 μg in 100 μL PBS per mouse) and an equal amount of free Cy7 (dye alone, negative control) were intravenously injected (i.v.) into C57/BL6 mice via the tail vein. At the indicated time points after injection, the mice were sacrificed by an overdose of pentobarbital sodium anesthesia, and the organs, including the hearts, lungs, livers, kidneys, and spleens, were collected and observed using an IVIS Spectrum optical imaging system (Perkin Elmer, Waltham, MA, USA). The intensity of fluorescence was quantified by Analyze 12.0 software (Perkin Elmer). To detect the distribution of EVs in organs, frozen sections of lung, liver, and spleen were made and costained with anti-CD68 (Abcam, ab201340) and DAPI (Sigma). Digital images of the stained sections were captured using a confocal laser scanning microscope (Nikon).
Ivis spectrum optical imaging system
Manufactured by PerkinElmer
Sourced in United States
The IVIS Spectrum is an optical imaging system designed for in vivo analysis of bioluminescent and fluorescent signals in small animal models. It provides quantitative data on biological processes and gene expression. The system utilizes highly sensitive charge-coupled device (CCD) cameras to capture real-time images and data.
Automatically generated - may contain errors
Lab products found in correlation
Living image software 4, by PerkinElmer (3 mentions)
Living image software, by PerkinElmer (2 mentions)
D luciferin potassium salt, by GoldBio (2 mentions)
Analyze 12, by PerkinElmer (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Laser scanning confocal microscope, by Nikon (1 mentions)
D luciferin, by GoldBio (1 mentions)
D luciferin, by PerkinElmer (1 mentions)
Ellman s reagent, by Thermo Fisher Scientific (1 mentions)
Ivis living imaging software, by PerkinElmer (1 mentions)
18 protocols using ivis spectrum optical imaging system
1
In Vivo Biodistribution of EVs
2
Bioluminescence Imaging for Tumor Burden
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For tumor burden monitoring, mice were injected 3 mg D-Luciferin (Goldbio, St. Louis, MO, USA) i.p. for bioluminescence imaging (BLI) following a 5 min uptake period. An IVIS Spectrum Optical Imaging system (Perkin Elmer) was used to acquire BLI scans for 60 s or 1 s and signals were normalized for the luciferase activity [photons/second]. ROI scans were analyzed using Living Image Software 4 (Perkin Elmer), by drawing regions of interest (ROI) around the whole animal and corrected for background signal.
3
Leukemia Progression Monitoring in Mice
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All leukemia models were introduced by i.v. injection and transplanted into nonirradiated secondary recipient experimental animals. BALB/c mice were used for the WEHI-3B leukemia model (0.5 × 106/cells/mouse) and C57BL/6J mice for MLL/AF9-dsRed (0.2 × 106/cells/mouse). Leukemia progression was assessed by fluorescence (MLL/AF9 dsRed) using the IVIS-Spectrum Optical Imaging System (Caliper, PerkinElmer). Mice were shaved to reduce light attenuation.
4
Intranasal Delivery of Fluorescent LNPs
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Adult male and female nude mice were purchased from Charles River. The mice were maintained in the University of Manitoba Central Animal Care Facility under a temperature-controlled environment with a 12-hour dark-light cycle and unlimited access to food and water. All experiments complied with the Canadian Council on Animal Care guidelines and the protocol (#22-040) approved by the University of Manitoba Animal Care Committee. Before intranasal administration, the mice were anesthetized with 2.5% isoflurane. The LNP formulation (LNP-PEP and LNP-siACE2) in PBS was administered with a Hamilton microsyringe with a blunt needle tip inserted 3 mm into the nostril (5 µL per nostril; 4 mg/Kg final dose). Serial images of the fluorescent LNPs were taken at different time intervals (5, 10, 15, 30, 60, 120, 180, and 1440 minutes) using a PerkinElmer/Caliper IVIS Spectrum optical imaging system (Ex/Em: 745/800 nm; Binning Factor: 4 and Exposure Time: 10 seconds). The fluorescence activity at different time points in specified regions of interest (ROI) was estimated as Total Radiant Efficiency [p/s]/[µW/cm²] using PerkinElmer Living Image® software.
5
Bioluminescence Imaging for Tumor Burden
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Optical Imaging was performed using an IVIS Spectrum Optical Imaging system (Perkin Elmer). For weekly tumor burden monitoring, mice were injected 75 mg/kg XenoLightTM D-Luciferin (Perkin Elmer, Waltham, MA, USA) i.p. and luciferase activity was detected using bioluminescence imaging (BLI) following a 5-min uptake. Images were acquired using a 5-s exposure, f-Stop 8 and binning 8 with a field of view of C-14. Signals above a 10% threshold were normalized for the photon radiance [photons/second/cm2/sr] and quantified using Living Image Software 4 (Perkin Elmer).
6
Luminescence Assay for A431-Luc2 Cells
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Luminescence of A431-Luc2 cells (passage 7 and 11) was assessed prior to tumor challenge. Cells were seeded in duplicate in a black 96-well plate by 2-fold serial dilution starting with 40,000 cells. Cells were allowed to attach for 4 hours. A stock of D-luciferin potassium salt (Gold Biotechnology, Olivette, MO) 15 mg/ml was initially prepared, 200 µl of luciferin stock (15 mg/mL) were added to 10 ml of growth media, then 100 µl of this mixture were added to each well for a final concentration of 150 µg/ml. Luminescence was quantified as total flux (photons/second) using the IVIS spectrum optical imaging system and the Living Image® software 4.5.5 (Perkin Elmer, MA, USA). Data were analyzed using GraphPad Prism software v7.03 (Boston, MA).
7
In Vivo Cell Tracking using Near-Infrared Dye
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Near-infrared dye DiR was used to detect transplanted cells in vivo since it has high tissue penetration and a low false-positive signal4 (link),5 (link). Three million hiPSC-MSCs were stained with 5 μg/mL DiR cell-labeling solution (D12731, Life Technologies, Carlsbad, CA, USA) for 15 min at 37 °C, then washed and re-suspended in 30 μl fresh plain medium for intramuscular transplantation6 (link). To quantify and detect the biodistribution of transplanted cells, animals were prepared for epi-fluorescent imaging using an IVIS® Spectrum optical imaging system under field of view 3.9 × 3.9 cm (PerkinElmer, Inc. MA, USA). All the animals laid on the back. The engrafted DiR-labeled cells were visualized under a 750 nm/800 nm excitation/emission wavelength with fluorescent intensity represented by radiant efficiency. To estimate the survival rate of the transplanted cells, the averaged radiant efficiency was calculated. The estimated survival rates at day 14, 21, 28, or 35 was calculated as the percentage of the radiant efficiency at day 14, 21, 28, or 35 versus that of the averaged radiant efficiency just after intravenous transplantation.
8
In Vivo Biodistribution of Lipo-DiR
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SKOV-3 tumor-bearing BALB/c nude mice were separately and intravenously injected with 10 nmol mPEG × DNS-modified Lipo-DiR or mPEG × HER2-modified Lipo-DiR when the tumor size was 200 mm3. The fluorescent signal of Lipo-DiR was monitored by IVIS spectrum optical imaging system (excitation, 750 nm; emission, 780 nm; PerkinElmer, Waltham, MA, USA) at 24, 48 and 72 h post-injection. The tumors and different organs (liver, spleen, heart, lung, ovary and kidney) of each group were collected at 72 h after Lipo-DiR injection. As previous study described23 (link), the region of interest (ROI) in tumors or different organ areas were drawn and analyzed with Living Image software version 4.2 (Caliper Life Sciences).
9
Orthotopic Prostate Cancer Xenograft Model
10
Multimodal Imaging of Biological Samples
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CT imaging was carried out on a Micro CT (TriFoil Imaging CT120) with fast scan mode. MR imaging was performed on a MRI scanner (MR solution, 3T) using T2 sequence. Bioluminescence imaging was performed on an IVIS Spectrum optical imaging system (PerkinElmer, USA). MPI was imaged using a MPI scanner (Magnetic Insight Inc, MOMENTUM™ Imager). Photoacoustic imaging was conducted on a photoacoustic imaging system (Vevo LAZR, Visual Sonics Company, Canada). The detailed parameters for imaging are described in supporting information .
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