Ivis lumina xr system

Female athymic nude mice bearing a subcutaneously COLO-205 tumor (approximately 350~500 mm³) were intravenously injected with Cy5.5-labeled RGD-TRAIL and Cy5.5-RGD-TRAIL-ELP at a dosage of 5 mg/kg (100 μg). In vivo fluorescence imaging was performed by scanning the mouse abdomen at the predetermined time interval (2, 4, 8, 24 h) using IVIS Lumina XR system (Caliper, Life science) with Cy5.5 excitation (640 nm) and emission (672 nm) filter sets. At 24 h post injection, the mice were sacrificed. Their major organs and tumor were harvested. Each organ and tumor were rinsed with saline three times and put into the board. To analyze the organ distribution, all data are calculated using the region-of-interest (ROI) function of analysis workstation software and data were given as mean ± S.D. for a group of three animals. A quantification of fluorescence signals was performed as total photons per centimeter squared per steradian (p/s/cm²/sr) per each organs and tumor.

Animal experiments were performed following the protocols approved by Institutional Animal Care and Use Committees of Jilin University. Nude (BALB/C-nu/nu) mice and SCID (severe combined immunodeficient) mice were obtained from Vital River Laboratory Animal Technology (Beijing, China). 5×10⁶ PANC-1 cells stably transduced with lentivirus-based pGCSIL-GFP vector carrying negative control and RPL34-siRNA were suspended in HBSS and injected subcutaneously into the flank region of 6-week-old female athymic nude (BALB/C-nu/nu) mice (Vital River Laboratory Animal Technology, Beijing, China). The tumors were monitored and grown to an average volume of 200 mm³. Tumor size was assessed by caliper every other day and tumor volumes were calculated after 24 days using the formula: V = 4/3 × π (length/2 × (width/2)²).
To establish a metastasis model, SCID mice were injected with 1× 10⁶ viable PANC-1-luc-NC and PANC-1-luc-KD cells via tail veins. Successful injection was confirmed by immediate luciferase imaging. For luciferase imaging, mice were anesthetized with isoflurane and intraperitoneally injected with luciferin (25 mg/ml in 0.1 ml PBS). Animals were imaged 15 min after injection using an IVIS LuminaXR system (Caliper, Hopkinton, MA, USA) once a week for a total of six weeks.

The bio-distribution and tumor-targeting properties of CA IX-Lips in A549-Luc orthotopic lung cancer BALB/c nude mice were investigated using a IVIS Lumina XR system (Caliper LifeSciences). A CA IX-decorated DiR liposomes suspension was administered directly via pulmonary delivery using a Microsprayer Aerosolizer Pulmonary Aerosol Kit for Mouse Model PAK-MSA (Penn-Century, Inc. Wyndmoor, PA19038 USA), and the distribution behavior in the nude mouse was monitored at different time points after endotracheal administration using an in vivo imaging system.

Purified ANXA5-TagRFP (2 mg kg⁻¹) was injected into nude mice through the tail vein. After 30 min, in vivo imaging was visualized by an IVIS Lumina XR system (Caliper Life Sciences, Hopkinton, USA) with TagRFP excitation (570 nm) and emission (672 nm) filter sets. After dissecting mice, Ex vivo fluorescence of various tissues was photographed and analyzed for ANXA5-TagRFP. Similarly, BALB/c mice with TNBS-induced colitis were injected by the tail vein with ANXA5-TagRFP (2 mg kg⁻¹). After 30 min, mice were dissected to examine the fluorescence intensity of ANXA5-TagRFP in various tissues by an IVIS Lumina XR system and analyzed by Living Image software.

The imaging was performed with IVIS Lumina XR system (Caliper Life Sciences). To limit possible interference of melanin in B6 mouse skin with the signal generated by the luciferase activity, we used WT Balb/c mice for these experiments. Mice injected with PBS, mRNA-LNP coding for luciferase or PR8 HA, were intraperitoneally injected with D-Luciferin (Potassium Salt, Goldbio) at the dose of 150 mg/kg. Five minutes later, the mice were anesthetized in a chamber filled with 3% isoflurane for 1 minute, then transferred to the imaging platform with maintained 2% isoflurane via gas ports. With the Living Image Software provided by Caliper, the signal was acquired by measuring total flux (photons/sec) for 5 seconds exposure time. The total flux is the radiance (photons/sec/cm²/steradian) in each pixel summed or integrated over the region of interest (ROI) area (cm²) x 4π. The total flux values were normalized to ensure accurate quantitation and comparability. Briefly, the normalized value (Vn) in each independent experiment was calculated as following:
$$\mathrm{V}\mathrm{n}=\mathrm{V}\mathrm{o}÷\overline{\mathrm{V}\mathrm{s}}$$
Vo was original total flux value of a given sample at a given timepoint; $\overline{\mathrm{V}\mathrm{s}}$ was the mean of all values from all groups.

The imaging was performed with IVIS Lumina XR system (Caliper Life Sciences). To limit possible interference of melanin in B6 mouse skin with the signal generated by the luciferase activity, we used WT Balb/c mice for these experiments. Mice injected with PBS, mRNA-LNP coding for luciferase or PR8 HA, were intraperitoneally injected with D-Luciferin (Potassium Salt, Goldbio) at the dose of 150 mg/kg. Five minutes later, the mice were anesthetized in a chamber filled with 3 % isoflurane for 1 minute, then transferred to the imaging platform with maintained 2 % isoflurane via gas ports. With the Living Image Software provided by Caliper, the signal was acquired by measuring total flux (photons/sec) for 5 seconds exposure time. The total flux is the radiance (photons/sec/cm²/steradian) in each pixel summed or integrated over the region of interest (ROI) area (cm²) × 4π. The total flux values were normalized to ensure accurate quantitation and comparability. Briefly, the normalized value (Vn) in each independent experiment was calculated as following:
$$\text{Vn}=\text{Vo}÷\overline{\text{Vs}}$$
Vo was original total flux value of a given sample at a given timepoint; $\overline{\text{Vs}}$ was the mean of all values from all groups.