In vivo imaging was performed on the Pearl Trilogy Small Animal Imaging System (LI-COR Biosciences, Lincoln, NE) with wavelength excitement at 800 nm. MUC4-IR800 was administered via tail vein injection. A dose of 50 μg was chosen based on prior studies.15 (link),16 (link) The LS174T subcutaneous model was administered MUC4-IR800 (50 μg) and imaged at 48 and 72 hours. The subcutaneous C4 and Liver 2 models were administered MUC4-IR800 (50 μg) and imaged at 24, 48, and 72 hours. The orthotopic models all received MUC4-IR800 (50 μg) and were euthanized 48 hours later. A laparotomy was performed to expose intra-abdominal organs. The mice were then imaged on the Pearl Trilogy Small Animal Imaging System (LI-COR Biosciences, Lincoln, NE). The Pearl Trilogy Small Animal Imaging System Software (LI-COR Biosciences, Lincoln, NE) was used to quantify the strength of the MUC4-IR800 signal. For the subcutaneous models, skin was used as background, and a tumor to background ratio (TBR) was derived by dividing the tumor signal by the background signal. For the LS174T and C4 primary tumor PDOX models, normal bowel was used as the background signal to calculate the TBR. For the Liver 2 liver PDOX model, the normal liver was used as the background signal to calculate the TBR.
Pearl trilogy small animal imaging system
Manufactured by LI COR
Sourced in United States
The Pearl Trilogy Small Animal Imaging System is a multi-modal imaging platform designed for preclinical research. It combines fluorescence, bioluminescence, and X-ray imaging capabilities in a single compact system. The device enables non-invasive, longitudinal studies of small animal models.
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Lab products found in correlation
Irdye 800cw, by LI COR (4 mentions)
Psvue794 reagent kit, by LI COR (3 mentions)
Nude mice, by Charles River Laboratories (1 mentions)
Collagenase clostridium histolyticum type 4, by Merck Group (1 mentions)
Irdye800cw maleimide, by LI COR (1 mentions)
Fluoromax fluorometer, by Horiba (1 mentions)
Uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Mounting media, by Vector Laboratories (1 mentions)
Bx63 fluorescence microscope, by Olympus (1 mentions)
Image studio lite software, by LI COR (1 mentions)
Irdye 800cw 2 dg optical probe, by LI COR (1 mentions)
Evans blue, by Merck Group (1 mentions)
Image studio software version 5, by LI COR (1 mentions)
Image studio software, by LI COR (1 mentions)
Ivis spectrum in vivo imaging system, by PerkinElmer (1 mentions)
30 protocols using pearl trilogy small animal imaging system
1
In Vivo Imaging of MUC4 Expression
2
Multimodal Optical Imaging Systems
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The Pearl Trilogy Small Animal Imaging System (LI-COR Biosciences, NE, USA) allows white-light and near-infrared imaging at 700 and 800 nm in a black-box setting, minimizing the interference from ambient light. It is used as a gold standard for quantification and comparison of in vivo fluorescent signals. To demonstrate the potential of the dyes for clinical translation the mini-FLARE Imaging Systems Kit, FLARE Model R1 (both Curadel, Natick, MA, USA) and Artemis (Quest Medical Imaging, Netherlands) near-infrared camera systems were used. Each of these systems is equipped with 700 nm and 800 nm channels next to visual color, and is suitable for intraoperative use. Using mini-FLARE, excitation wavelengths for 700 nm and 800 nm NIR fluorophores were 665 ± 1 nm and 760 ± 1 nm, respectively, with typical fluence rates of 1–10 mW/cm2.
3
Quantitative in vivo and ex vivo Imaging
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In vivo and ex vivo optical images were acquired with a Pearl® Trilogy Small Animal Imaging System (LI-COR, Lincoln, NE). Delivery of the NPs was confirmed by imaging the mice in vivo at 48 h after the first injection and either at 24 h (group 1) or at 48 h (group 2) after the second injection. Mice were sacrificed and organs excised for ex vivo quantification. Excised tumors, kidneys, liver, spleen, heart, lungs, and muscle were imaged. Fluorescent intensities in regions of interest (ROIs) were quantified by using Living Image 4.5 software (Caliper, Hopkinton, MA). The tumors were sectioned into two to three slices of ~1 mm thickness. Fluorescent signal was acquired from both sides of each slice, and the values acquired for each tumor were averaged. Signal intensities were normalized to the area of the ROI.
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In vivo and ex vivo optical images were acquired with a Pearl® Trilogy Small Animal Imaging System (LI-COR, Lincoln, NE). Delivery of the NPs was confirmed by imaging the mice in vivo at 48 h after the first injection and either at 24 h (group 1) or at 48 h (group 2) after the second injection. Mice were sacrificed and organs excised for ex vivo quantification. Excised tumors, kidneys, liver, spleen, heart, lungs, and muscle were imaged. Fluorescent intensities in regions of interest (ROIs) were quantified by using Living Image 4.5 software (Caliper, Hopkinton, MA). The tumors were sectioned into two to three slices of ~1 mm thickness. Fluorescent signal was acquired from both sides of each slice, and the values acquired for each tumor were averaged. Signal intensities were normalized to the area of the ROI.
4
Tumor Targeting with Labeled Antibodies
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All animal studies were performed per the guidelines of the IACUC and with protocols approved by the Washington University Division of Comparative Medicine. The TAF15 antibody or isotype control antibody was labeled with IRDye 800CW as per manufacturer’s instructions (Licor). Cancers were induced by injecting A549 or H460 cells in the right hind limbs of nude mice (Charles River). The cancers were irradiated with three fractions of 3Gy or 0Gy (sham) over the course of 24 h. The cancer-bearing mice were then injected with 10 μg of labeled antibodies via the tail vein. Mice were imaged using the Pearl Trilogy small animal imaging system (Licor). Fluorescence was detected using an 800 nm channel. Images were analyzed using the Image Studio software. Background subtracted signal intensity was plotted using Graph Pad Prism software.
5
Quantitative Analysis of S. aureus Infection
6
Non-invasive Hydrogel Degradation Monitoring
7
Optimized Imaging of Resected Specimens
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To acquire images, the resected specimens were collected directly after surgery and transported to the Department of Pathology. Upon arrival, ink was applied to the resection plane by the pathologist to maintain the correct orientation of the resection plane with respect to the tumor. Directly after inking or after fixation of the specimen with formalin, the specimens were cut into ±5 mm thick tissue slices (i.e., bread loaves). The bread loaves were imaged on both sides in the PEARL Trilogy Small Animal Imaging System (LI-COR Biotechnology, Lincoln, Nebraska, United States). The PEARL imaging system standardizes factors affecting the measured fluorescence intensity (i.e., ambient light, the distance between the camera and imaged tissue, and the angle between the incoming light and the imaged surface) and is suitable for imaging fluorescent dyes around 700 and 800 nm.38 (link)– 40 (link, link) The parameters exposure and acquisition time are automatically set by the system, but it does not auto-gain the fluorescence intensities.39 (link) A white light and a fluorescence image are both sequentially acquired while imaging with the system. The focus settings were adjusted so that the camera was focused on the specimen. The resolution was fixed at , resulting in images of .
8
Dynamic CSF Flow Imaging in Rats
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Rats were anesthetized, placed in a stereotactic frame and through a burr hole in the lateral ventricle (same coordinates as for ICP and ventriculo-cisternal perfusion) a Hamilton syringe (RN 0.40, G27, a20, Agntho’s) was placed (4 mm deep) with 15 µl -aCSF with 10 µM carboxylate dye (MW = 1,091 Da, IRDye 800 CW, P/N 929–08972, LI-COR Biosciences). Immediately after the dye injection, the rat was swiftly placed in a Pearl Trilogy Small Animal Imaging System (LI-COR Biosciences) and within 1 min after ventricular dye injection, images were obtained at 30 s intervals (800 nm channel, 85 μm resolution, for 5 min). A white field image was acquired at the termination of each experiment, after which the rat was sacrificed. The isolated brain was then bisected to expose the ventricles to record a final micrograph ensuring proper targeting of the ventricular compartment. Images were analyzed in a blinded fashion using LI-COR Image Studio 5.2 (LI-COR Biosciences) and data presented as fluorescence intensity in a region of interest placed in line with lambda, normalized to the signal obtained in the first image. Five rats in each group were employed for the live-imaging of CSF flow.
9
In vivo and ex vivo Optical Imaging of COX-2 siRNA Nanoplex
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In vivo optical images were acquired with a Pearl® Trilogy Small Animal Imaging System (LI-COR, Lincoln, NE), ex vivo optical images were acquired with an IVIS Lumina Series III Spectrum scanner (Perkin-Elmer, Waltham, MA), and fluorescence intensities in regions of interest (ROIs) were quantified by using Living Image 4.5 software (Caliper, Hopkinton, MA). For in vivo optical imaging of the distribution of COX-2 siRNA/compound 4 nanoplex, MDA-MB-231 tumor bearing mice were injected intravenously with 100 µL of dextran conjugated COX-2 siRNA nanoplex (dextran 2.0 mg/mouse, N/P=20. COX-2 siRNA, 4 nmol/mouse) through the tail vein. Delivery of the nanoplex was confirmed by imaging the mice at 24 h and 48 h. Subgroups of mice were sacrificed either at 24 h or at 48 h after nanoplex injection for ex vivo imaging studies, and tumors and muscle were excised to obtain the optical images.
10
Exosome Labeling and Biodistribution
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Exosomes (approximately 1 μg/μl protein level) were treated for 30 min with 1 mM DiR (Invitrogen D12731) in a volume ratio of 500:1. Free dyes were cleared by centrifugation. DiR-labeled exosomes (100 μg at protein level) were injected into mice with indicated treatments via tail veins. Exosome localization in the body and individual organs was detected by Pearl Trilogy Small Animal Imaging System (LI-COR, United States) 12 h after injection.
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