Fluorescence imaging was performed using a multimodal IVIS Lumina XR imaging system (PerkinElmer). The mice were euthanised by cervical dislocation, followed by exposure of the abdominal cavity and removal of the intestine from the stomach to the rectum [33 (link)]. The instrument background fluorescence was removed using an adaptive fluorescence background subtraction tool. Images were analysed using Living Image version 4.3.1 [33 (link)].
Ivis lumina xr
Manufactured by PerkinElmer
Sourced in United States, Japan
The IVIS Lumina XR is a preclinical in vivo imaging system developed by PerkinElmer. It is designed to enable non-invasive, real-time visualization and quantification of bioluminescent and fluorescent signals from small animal models. The system utilizes highly sensitive optical and imaging technologies to capture detailed images and data from live animals.
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Lab products found in correlation
D luciferin, by PerkinElmer (4 mentions)
Living image software, by PerkinElmer (3 mentions)
Vivoglo, by Promega (2 mentions)
Isoflurane, by RWD Life Science (1 mentions)
Quantum gx2, by PerkinElmer (1 mentions)
Stereomicroscope, by Leica (1 mentions)
Prism 6, by GraphPad (1 mentions)
Living image software version 4, by PerkinElmer (1 mentions)
Nod scid gamma mice, by Jackson ImmunoResearch (1 mentions)
Basic fibroblast growth factor (bfgf), by Fujifilm (1 mentions)
Basic fibroblast growth factor (bfgf), by PerkinElmer (1 mentions)
D luciferin potassium salt, by GoldBio (1 mentions)
D luciferin, by Thermo Fisher Scientific (1 mentions)
Malachite green phosphate assay kit, by Merck Group (1 mentions)
6 mercapto 1 hexanol, by Merck Group (1 mentions)
Living image v 4, by PerkinElmer (1 mentions)
Prosense 750, by PerkinElmer (1 mentions)
Xenolight d luciferin, by PerkinElmer (1 mentions)
Living image analysis software, by PerkinElmer (1 mentions)
Amicon ultracentrifugation filter units, by Merck Group (1 mentions)
38 protocols using ivis lumina xr
1
Fluorescence Imaging of Intestinal Tissue
2
Targeted Atherosclerotic Plaque Imaging with SR-A-Ce6NB
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Ttargeted uptake of SR-A-Ce6NB into the atherosclerotic plaque was assessed by fluorescence imaging. Plaque-bearing mice were divided into targeted and non-targeted groups (n = 3 per group). For the targeted group, mice were intravenously administered SR-A-Ce6NB (80 μg/ mL, 0.2 mL/10 g); for the non-targeted group, mice were injected with Ce6NB or Ce6 (80 μg/mL, 0.2 mL/10 g) via the tail vein. After anesthesia with isoflurane (RWD Life Science, Shenzhen, China), mice in all groups were sacrificed at 0.5 h post injection and aortas were harvested for ex vivo fluorescence imaging using a fluorescence imaging system (IVIS Lumina XR, PerkinElmer Health Sciences, USA). Quantitative analysis was performed using living image software 4.4 for IVIS Lumina XR. Additionally, the aortic plaques were dissected at 0.5 h after injection of SR-A-Ce6NB for frozen sections to observe the accumulation of SR-A-Ce6NB within plaque using CLSM.
3
Femur Bone Density Analysis in Mice
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Mice were inspected with a fixation X-ray system (IVIS Lumina XR, PerkinElmer, Waltham, MA, USA), and the femur’s bone mineral density (BMD) was determined by a dual-energy X-ray absorptiometry system. Scans of uncalcified specimens were carried out using X-ray microtomography (micro-CT, Quantum GX2, PerkinElmer, Waltham, MA, USA) at 50 kV, 80 µA, 0.5-mm aluminum filtering, and 9-µm isotropic resolution. Each specimen was scanned for a total of 4 min. Using Analyze 14.0 software (USA), the bone tissue 1 mm from the end of the distal femoral growth plate with a layer thickness of 2 mm was selected as the region of interest (ROI) for 3D reconstruction of bone trabeculae. The bone tissue 3 mm from the distal femoral growth plate and extending 1 mm in thickness toward the backbone was selected as the ROI for cortical bone reconstruction. The image information was extracted with a minimum threshold of 150, and several parameters were quantitatively analyzed with the Analyze 14.0 software, including the BMD, cortex, intra-trabecular tissue, and trabeculae.
4
Orthotopic Transplantation of Transfected Mammary Epithelial Cells
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The transfected MECs under culture were treated with dispase for 1–2 h at 37 °C and then approximately 3% of all transfected cells were suspended in 4–10 μL of DMEM/F12 including 10% Matrigel per transplantation site and transplanted into the cleared fat pads of the inguinal mammary glands of rag2−/− mice from which the endogenous epithelium had been removed using a 50-μL syringe equipped with a 30-G needle (ITO, Shizuoka, Japan). Mammary repopulation was analyzed by detecting bioluminescence using an in-vivo imaging system (IVIS Lumina XR, PerkinElmer, Waltham, MA, USA) and mCherry fluorescence using a stereomicroscope (Leica, Wetzlar, Germany).
5
In Vivo Bioluminescence and Fluorescence Imaging
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IVIS Lumina XR (PerkinElmer, USA) was carried out for in vivo FLI and BLI imaging of 4T1 and U87MG tumor bearing BALB/c nu mice. OD‐TH (2.5 mg kg−1 Dox) was i.v. injected into mice when the tumor grew to about 200 mm3. After isoflurane anesthesia, the images were recorded from 1 to 48 h (Ex = 500 nm/Em reference DsRed). The solid tumor and major organs were excised 48 h later for FLI. In semi‐quantitative FLI analysis, DOX (2.5 mg kg−1) and OD‐TH (2.5 mg kg−1 Dox) were i.v. injected into different 4T1 tumor bearing mice, respectively. 24 later, their solid tumors and main organs were obtained. PBS solution (10 µL g−1) D‐fluorescein potassium salt (15 mg mL−1) was i.p. injected into U87MG tumor bearing BALB/c nu mice. The BLI of the mice head was measured 12 min post injection. The same region‐of‐interest (ROI) was circled by Living Image software (PerkinElmer, USA) after Flat Field Correction and Cosmic Ray Corrections to compare the FL intensity of parallel parts.
6
Non-invasive Optical Bioluminescence Imaging
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After MRI, the animals were placed in a dark chamber of the IVIS Lumina XR optical imager (Perkin Elmer, USA) in order to detect bioluminescence signals. Firstly, photographs were taken for anatomical coregistration of the bioluminescent source. Secondly, optical images were measured before and after intravenous administration of D-Luciferin dissolved in sterile PBS (50 mg/kg of body weight) with an exposure time of 1 minute, open aperture, and open emission filter. The images were acquired from a time series lasting 14 minutes, while the area under the dynamic time curve was calculated using GraphPad Prism 6.02 (GraphPad Software Inc, USA) in order to minimise the variability of D-Luciferin administration among the measurements. The bioluminescence examination was performed at the same time points as the MRI experiments until day 120 after PIs transplantation.
7
Lung Fluorescence Imaging in Mice
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Mice were divided into three groups. The first group was administered PBS intranasally as a negative control (n = 3). The second group was administered nCA intranasally as a positive control (n = 3). The third group was administered nCA on day 0 and injected with PEG-fDAO (10 U/mL, 0.1 mL/mouse) (n = 3) on day 9, followed by intraperitoneal injection of d -proline (1 M, 0.5 mL/mouse) on days 10, 11, and 12. Rhodamine-BSA (10 mg/kg) was injected one day before sacrifice on day 21. Fluorescence imaging of the excised lungs was performed using an IVIS Lumina XR (PerkinElmer Japan Co., Ltd.).
8
Therapeutic Evaluation of PEG-fDAO in Mice
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Mice were divided into three groups. The first group was administered PBS intranasally as a negative control (n = 3). The second group was administered nCA intranasally as a positive control (n = 3). The third group was administered nCA on day 0 and injected with PEG-fDAO (10 U/mL, 0.1 mL/mouse) (n = 3) through the tail vein on day 4, followed by intraperitoneal injection of d -phenylalanine (0.1 M, 0.5 mL/mouse) on days 5, 6, and 7 instead of d -proline. Rhodamine-labeled bovine serum albumin (rhodamine-BSA) (10 mg/kg) was injected 1 day before sacrifice on the 14th day. Fluorescence imaging of the excised lungs was performed using the IVIS Lumina XR (excitation: 555–585 emission: 695–770 nm; PerkinElmer Japan Co., Ltd., Kanagawa, Japan).
9
In Vivo Biocompatibility of Hydrogels
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All animal experiments performed for this study were carried out in compliance with the ARRIVE guidelines and the protocol was approved by the Committee on the Ethics of Animal Experiments of National Defense Medical College (approved numbers: 19010 and 19065). All methods were performed in accordance with relevant guidelines and regulations of National Defense Medical College. Female ICR mice (~ 30 g) were used for in vivo biocompatibility studies. A 1 cm incision was made in the mediodorsal skin of the mice, and a lateral subcutaneous pocket was prepared. Hydrogel samples (10 × 1 mm cylinders) were implanted under sterile conditions. At designated time intervals (days 1, 3, 7, and 14), mice (n = 5 for each day) were sacrificed, and the fluorescence of the remaining hydrogel in mice subcutaneous tissue was imaged using the IVIS Lumina XR (Ex: 460 nm, Em: 620 nm), and the intensity of fluorescence was analyzed using Living Image Software (PerkinElmer Inc., MA) and were processed for histological analyses.
10
In vivo Tumor Imaging Using Aptamers
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Mice bearing CRC119x tumors in the right flank were injected via tail vein with 2 nmol AF750-E3 or AF750-C36 and imaged at different time points up to 48 h post-aptamer injection on an IVIS Lumina XR (PerkinElmer, Waltham, MA, USA).
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