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Pe ivis spectrum

Manufactured by PerkinElmer
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The PE IVIS Spectrum is an in vivo imaging system designed for small animal research. It provides high-sensitivity detection and quantification of bioluminescent and fluorescent signals within living subjects.

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7 protocols using pe ivis spectrum

1

In vivo Tumor Suppression via M1 Macrophage Hydrogel

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Male athymic nude mice (BALB/c nu/nu, 4-6 weeks old) were used for the subcutaneous tumor animal model. Surgical procedures were as described previously 12 (link). Briefly, 2x106 MHCC97L cells tagged with luciferase, suspended in 0.2mL DMEM, were injected subcutaneously into the flanks of the mice. After 3 weeks of injection as the size of tumor reached approximately 3mm, the mice were divided into 3 groups (Negative control: DMEM; Blank: hydrogel with PBS; M1 gel: hydrogel embedded with 1x106 M1 macrophages; each group with 4 mice). 200μL of hydrogel was injected directly adjacent to tumor nodule weekly in the blank and M1 hydrogel group. Followed by the treatment, the tumor size of MHCC97L xenograft was monitored weekly by PE IVIS Spectrum in vivo imaging system (PerkinElmer). All mice were euthanized at week 4 and the size of tumor was measured and compared.
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2

Biodistribution and T3 Levels of Lipid Nanoparticles

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8-week-old male C57BL/6 N mice were administered with free Cy5 (FCy5), LCy5, and PLCy5 at a Cy5 dose of 1 μmol/kg by intraperitoneal injection. The mice were sacrificed at various time points after injection. iWAT, eWAT, iBAT, liver, spleen, heart, lung, kidney, brain, and soleus were collected and washed three times with Hank’s Buffered Salt Solution. The fluorescence intensity (Ex 650 nm, Em 670 nm) of tissue homogenates was quantified with a plate reader (BMG LABTECH, model 430-101). The ex vivo fluorescence signal was detected by PE IVIS Spectrum (PerkinElmer, model 124262).
For measurement of T3 concentrations in different tissues, 8-week-old male C57BL/6 N mice were injected with saline, FT3, LT3, or PLT3 at a dose of 2 μg per mouse. The mice were sacrificed at 8 h after injection. iWAT, eWAT, iBAT, liver, spleen, heart, lung, kidney, brain, and soleus were collected and homogenized. T3 concentrations in tissue homogenates were quantified with a commercial ELISA kit (Invitrogen, USA). Results were presented as fold change of T3 level relative to endogenous T3 level in each tissue of saline-treated mice.
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3

Bioluminescent S. aureus Antibiotic Assay

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A single colony of bioluminescent S. aureus strains containing plasmid pGLhla (Table 2) from BHI agar was resuspended in 200 μL sterile water, diluted to 75 mL 0.7% (w/v) soft agar, and plated on BHI agar. On the overlay, antibiotic discs (diameter 6 mm; Advantec Co., Tokyo, Japan) were placed, and the plates were incubated at 37 °C. After 20 h, inhibitory zones were determined, and luminescence was detected using a PE IVIS Spectrum in vivo imaging system (PerkinElmer, Hong Kong, China) [6 (link)]. When combining antibiotics and compounds, antibiotics (10 mM, 5 μL) and M21 (50 mM, 5 μL) were placed on the plates at a distance of 1.5 cm and 2 cm, respectively. When analyzing the interaction between cefoxitin and M21, we used a concentration of 4 mM of antibiotic and M21 and the distance between the discs was 1.5 cm.
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4

Bioluminescence-based Cell Imaging

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Luciferase-labeled cells were embedded in Matrigel (Corning Inc., Corning, NY) and quantified by bioluminescence-based live-cell imaging using PE IVIS Spectrum in vivo imaging system (PerkinElmer, Inc., Waltham, MA). Enduren (Promega Corporation, Madison, WI) was used as the substrate for Renilla luciferase.
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5

Assessing miR-509-3p Tumor Suppression

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HCT116 cells stably transduced with miR-509-3p (pre-mir) or empty vector were transfected with a Firefly luciferase expressing plasmid and selected by puromycin. For the animal experiments, Luc-HCT116 cells (5,000,000) stably transduced with miR-509-3p (pre-mir) (N = 7) or empty vector (N = 4) were injected orthotopically into the cecal wall of the NOD/SCID mice. Sample size used per study arm were selected randomly to achieve a basic statistical power for evaluation. After ~ 4 weeks, the mice were euthanized by rendering them unconscious first using Sodium Pentobarbital (40–60 mg/kg IP) followed by cervical dislocation. They were subsequently dissected, and tumor development was observed by capturing the bioluminescence signal using the PE IVIS Spectrum in vivo imaging system (PerkinElmer).
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6

Evaluating Nanoparticle-Enhanced Anti-Tumor Therapy

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Animal experiments were approved by the Ethics Committee of Weifang Medical University (2020SDL117). 5 × 106 MDA-MB-231 cells were subcutaneously injected into the right hind limb of BALB/c nude female mice (age: 4 weeks; weight: 12–17 g; Vital River, Beijing, China). When tumor volume reached to 70–130 mm3, 30 mice were randomly divided into 6 groups: control (PBS), lapatinib (15 mg/kg), PAB (10 mg/kg), combination (lapatinib/PAB), vehicle (ferritin) and nanoparticle (L/P@Ferritin). Each group was treated with drugs by intraperitoneal injection every 2 days for seven times. Tumor sizes were evaluated and calculated by the following formula: 0.5 × length × width2. The mice were typically euthanized by CO2, tumors, and organs were collected for immunofluorescence and HE stain.
To generate lung metastases model, 1 × 106 MDA-MB-231-luc cells were injected intravenously into NOD-SCID immunodeficient mice (female, age: 4 weeks; Vital River, Beijing, China). 12 mice were randomly divided into three groups: control (PBS), combination (lapatinib/PAB) and nanoparticle (L/P@Ferritin). Each group was treated with drugs by tail vein injection every 3 days for seven times after inoculation 14 days. Lung metastasis was evaluated using the PE IVIS Spectrum (PerkinElmer, MA, USA).
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7

In Vivo Biodistribution of PEG-Coated Nanoparticles

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We chose 6–8 weeks old female Balb/c nude mice as animal model. The animal experiment had been approved by the Ethics Committee of Jiangyin People’s Hospital, the Jiangyin Clinical College of Xuzhou Medical University. To investigate the biodistribution of PEG-TE10-PLGA NPs in vivo, we injected 100 μL PBS containing 5 × 106 TE10/DOX cells into the subcutaneous breast. When the tumor grew to nearly 300 mm3, PLGA@DiR, TE10-PLGA@DiR, and PEG-TE10-PLGA@DIR (5 mg/kg) were injected through the tail vein, and the control group was injected with equal volume of saline. After 0, 8, 24, and 48 h, tail vein blood (200 μL) was taken and the serum was collected by centrifugation. The drug concentration in blood was evaluated through the fluorescence intensity of DiR. The fluorescence intensity at the tumor site was measured with a live animal imager (Perkin Elmer PE IVIS SPECTRUM, United States). After 48 h of intravenous injection, the mice were dissected, heart, liver, spleen, lung, kidney and tumor were taken out and the fluorescence intensity in each organ and tumor tissue was measured.
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