To obtain noninvasive measurement of bacterial burden, mice were anesthetized with inhalation isoflurane (2%), the dorsal region of the mouse was shaved, and in vivo bioluminescence imaging was performed using the Xenogen IVIS Lumina II (PerkinElmer, Hopkinton, MA) on post-operative days (POD) 0, 1, 3, 5, 7, 10, 14, 17, 21, 25, 28, 32, and 35 as previously described.[26 (link),37 (link)–39 (link)] The Xen36 bioluminescent signals were obtained with a 5-minute acquisition time, a 13 cm field of view (FOV), and medium binning settings. Data are presented on a color scale overlaid on a grayscale photograph of mice and quantified as total flux (photons per second (s) per cm2 per steradian (sr) [photons/s/cm2/sr]) within a circular region of interest (1X103 pixels) using Living Image software (PerkinElmer, Hopkinton, MA).
Xenogen ivis lumina 2
Manufactured by PerkinElmer
Sourced in Switzerland
The Xenogen IVIS Lumina II is a high-performance in vivo imaging system designed for preclinical research. It uses bioluminescence and fluorescence imaging techniques to visualize and quantify biological processes in small animal models.
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4 protocols using xenogen ivis lumina 2
1
Noninvasive Bioluminescence Imaging for Bacterial Burden
2
Measuring Tumor Burden in Mice
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ID8 tumor cells (5 × 106/mouse) expressing luciferase were injected i.p. into 8–10-week-old C57/BL6 female mice as described earlier. Day 19 post-tumor implantation and before the first treatment, tumor burden was measured by bioluminescence quantification of luciferase activity (reported as photons/sec). Bioluminescence imaging (Xenogen IVIS lumina II, PerkinElmer, Switzerland) was performed once every 2 weeks thereafter before the onset of ascites formation in PBS-treated control mice.
3
Monitoring Tumor Burden and Progression
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B16F10 and LLC1 tumor burden was monitored with a digital caliper. Mice were euthanized when tumor burden reached 1cm3 or at the set endpoint (on day 21 post tumor implantation) for immune analysis. Mice harboring the ID8 cancer cells were monitored following animal weight (measured every 2 days until body weight exceeded >20% or when they became distressed and moribund); and bioluminescence quantification (Xenogen IVIS lumina II, PerkinElmer, Switzerland) of luciferase activity (reported as photons/sec). Bioluminescence imaging started 19 days post-tumor implantation and before the first treatment, then it was performed once every 2 weeks thereafter before the onset of ascites formation in PBS-treated control mice.
4
Systemic Delivery of miR-449a for Cancer Metastasis Inhibition
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The miR-449a oligonucleotide or miR-CON (GenePharma, Shanghai, China) was formulated with a polymer-based agent (in vivo-jectPEI; Polyplus, Illkirch, France) according to the manufacturer's instructions.17 (link) In the experiment involving systemic miR-449a treatment, the miR-449a or miR-CON oligonucleotide was formulated with a polymer-based agent (in vivo-jectPEI; Polyplus, Illkirch, France) according to the manufacturer's instructions and then intravenously injected into mice biweekly. miR-449a treatment was given five times in the multiorgan metastasis model, and three times in the lung metastasis model. Ten weeks after cell injection in the multiorgan metastasis model and six weeks after cell injection in the lung metastasis model, bioluminescent imaging was performed to observe the metastasis of cancer cells (Xenogen IVIS lumina II, PerkinElmer, MA). The signal was analyzed using Living Image R Software Version3.1.
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