The IVIS spectrum live imaging system (Perkin Elmer, Waltham, MA, USA) was used to assess the skin retention of drugs. Prior to the experiment, SD rats were anesthetized by 5% chloral hydrate, and the abdominal hair was depilated. Dissolving MNs loaded with both rhodamine B (20 μg) and propranolol hydrochloride (1.5 mg) were fabricated as described above, which were applied to the abdominal skin in SD rats for 10 min. For comparison, a rhodamine B solution (0.1%, 20 μL) was subcutaneously injected in the abdomen of an SD rat. The SD rats were imaged by IVIS at 30 min, 1, 2, 4, 6, 8, 10, 12, and 24 h after administration under Epi-Illumination mode with an excitation wavelength of 554 nm and emission wavelength of 580 nm.
Ivis spectrum live imaging system
Manufactured by PerkinElmer
Sourced in United States
The IVIS Spectrum Live Imaging System is a bioluminescent and fluorescent in vivo imaging platform. The system is designed to capture real-time images of living subjects, enabling non-invasive, longitudinal monitoring of biological processes.
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9 protocols using ivis spectrum live imaging system
1
Skin Retention of Dissolving MNs
2
Intracranial Xenograft Mouse Model for TMEM158
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All animal experiments were approved by the Ethical Committee of the Tianjin Medical University General Hospital. An intracranial xenograft mouse model was established as previously described [31 (link)]. U251MG cells were stably transfected with sh-TMEM158 and OE-TMEM158 lentivirus, and control cells were injected into the brains of mice. Tumor burden was monitored by bioluminescence imaging every week starting on day 7 after implantation using an IVIS Spectrum Live Imaging System (Perkin Elmer, USA). Body weight and OS of mice in both groups were monitored every day. The brains of the mice were carefully extracted when the mice died, fixed in 10% formalin, and embedded in paraffin for H&E staining and IHC staining.
3
Tumor Metastasis Tracking in Mice
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Male BLAB/C nude mice (Vital River, Beijing, China) aged 4–5 weeks were divided into 4 groups with 6 mice per group. 1.5 × 106 luciferase-expressing cells suspended in 100 µL PBS were intravenously in the tail vein into each mouse. After 4 weeks of cell injection, an IVIS Spectrum live imaging system (PerkinElmer, USA) was used to track the progress and metastasis of the tumor. Then the lungs were photographed and stained with HE after the mice were euthanized.
4
Evaluating PP and TMZ for GBM Treatment
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One week after tumor cells inoculation, the mice were randomized as four groups, including control (vehicle), intraperitoneal injection of PP at 0.5 mg/kg/day for 3 consecutive weeks, gavage of TMZ at 20 mg/kg/day for 3 consecutive weeks, and 0.5 mg/kg PP combined with 20 mg/kg TMZ (n = 5 mice per group). BLI was carried out to detect the intracranial tumor growth with IVIS Spectrum Live Imaging System (PerkinElmer, Branford, USA). The survival rate of mice in every group was observed.
In order to explore the mechanisms of PP on GBM in vivo, U87MG-Luc tumor-bearing mice were divided into two groups and intraperitoneally injected of PP at 0.5 mg/kg/day, or PBS for 3 consecutive weeks (n = 3 mice per group). All mice from the two groups were sacrificed 48 h after the last treatment (at day 30 after tumor implantation), and the brains or tumors of the mice in both groups were harvested for histopathology, TUNEL staining and Western blot analysis.
In order to explore the mechanisms of PP on GBM in vivo, U87MG-Luc tumor-bearing mice were divided into two groups and intraperitoneally injected of PP at 0.5 mg/kg/day, or PBS for 3 consecutive weeks (n = 3 mice per group). All mice from the two groups were sacrificed 48 h after the last treatment (at day 30 after tumor implantation), and the brains or tumors of the mice in both groups were harvested for histopathology, TUNEL staining and Western blot analysis.
5
Tracking hADSC Engraftment and Retention
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To evaluate hADSC engraftment and retention after treatment, hADSCs were labeled with VivoTrack 680 (PerkinElmer) or DiI (Thermo Fisher Scientific) and injected or delivered with CLP at the wound site. Luminescence intensity was monitored (the images were taken after Days 0, 1, 2, 3, 4, 9, and 14 days after surgery and each treatment) and quantified for 14 days using an IVIS Spectrum Live Imaging System (PerkinElmer).
6
Evaluating AZD8055 and Rapamycin in U87 Xenograft
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To evaluate whether AZD8055 and rapamycin inhibits tumor growth in vivo, the U87 neurospheres was selected to establish a nude mouse model. Female BALB/c nude mice (4–5 weeks, 15–17 g) were purchased from the Animal Center of the Cancer Institute at Chinese Academy of Medical Science (Beijing, China). To establish an intracranial model, 5 × 104 U87 initiating cells with a Luciferase-encoding lentivirus were injected into the mice (n = 5 per group) stereotactically. The mice were divided into three groups randomly. Luciferase-encoding lentivirus were purchased from GeneChem (Shanghai, China). The lentivirus vector is GV260: Ubi-MCS-Luc-IRES-Puromycin. The drugs were dissolved in 20%DMSO, 40PEG-300, and 40% PBS. After 7 days post implantation, the mice were injected intraperitoneally with AZD8055 (10 mg/kg/day), rapamycin (1.5 mg/kg/day) or DMSO 5 days a week during the survival period. Intracranial tumor growth was detected by using BLI on days 7, 14, and 28, using the IVIS Spectrum Live Imaging System (PerkinElmer, Branford, USA). The animal research was performed according to the internationally recognized guidelines and national regulations. Brains were extracted and fixed in 10% formalin and then, embedded in paraffin for HE.
7
Characterization of Nanoparticle Properties
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The physicochemical properties of various NPs were characterised after 20-fold dilution with purified water. The hydrodynamic diameter and zeta potential were determined using a Zetasizer Nano (Malvern Instruments, Malvern, Liverpool, UK) at 25 °C. The particle size was reported as the Z-average value. Samples (100 μL) were added to a 96-well plate and fluorescently quantified using an IVIS®spectrum live imaging system (PerkinElmer, CA, USA) with excitation/emission wavelengths of 710/760 nm.
8
Quantification of mPEG-PCL Nanoparticles
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Quantification of mPEG-PCL nanoparticles in both the blood and various tissues was attained by establishing linearity between particle-bound fluorescence intensity and the concentration of nanoparticles. The concentration of mPEG-PCL nanoparticles, which is expressed as the concentration of the constituting materials, was calculated based on calibration curves.
Blood was collected into tubes pretreated with heparin from the eye sockets of male SD rats. The heart, liver, spleen, lungs and kidneys were dissected, washed, weighted and homogenized with five volumes of physiological saline after sacrificing BALB/c mice by cervical dislocation. The suspension of mPEG-PCL nanoparticles was mixed with the blood or tissue homogenates to obtain calibration samples at a series of concentrations (0.0048, 0.0098, 0.02, 0.04, 0.078, 0.156, 0.312, 0.625, 1.25, and 2.50 mg/mL). The fluorescence intensity of 100 μL samples was measured by IVIS Spectrum Live Imaging System (PerkinElmer, Waltham, MA, USA)31 (link),35 (link). The calibration curve was obtained by linear regression of fluorescence intensity vs. nanoparticle concentration. The quantification method was validated following protocols utilized for evaluation of conventional analytical methods.
Blood was collected into tubes pretreated with heparin from the eye sockets of male SD rats. The heart, liver, spleen, lungs and kidneys were dissected, washed, weighted and homogenized with five volumes of physiological saline after sacrificing BALB/c mice by cervical dislocation. The suspension of mPEG-PCL nanoparticles was mixed with the blood or tissue homogenates to obtain calibration samples at a series of concentrations (0.0048, 0.0098, 0.02, 0.04, 0.078, 0.156, 0.312, 0.625, 1.25, and 2.50 mg/mL). The fluorescence intensity of 100 μL samples was measured by IVIS Spectrum Live Imaging System (PerkinElmer, Waltham, MA, USA)31 (link),35 (link). The calibration curve was obtained by linear regression of fluorescence intensity vs. nanoparticle concentration. The quantification method was validated following protocols utilized for evaluation of conventional analytical methods.
9
Intracranial Xenograft Mouse Model for U87 Glioma
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Experiments on mouse were approved by the Ethical Committee of Tianjin Medical University General Hospital. Intracranial xenograft mouse model was established as previously described.36 , 37 U87 cells stably transfected with shControl or shIKBKE lentivirus were injected into mice respectively. Bioluminescence imaging was performed to detect tumoral growth at seventh day after implantation through IVIS Spectrum Live Imaging System (Perkin Elmer, USA).
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