5×106 OVCAR-8-RFP cells suspended in PBS were injected intraperitoneally into NCr nu/nu athymic female mice (N=5) aged 10 to 12 weeks (Taconic, Rensselaer, NY). Tumor burden was monitored on a weekly basis using a Xenogen IVIS® Spectrum In Vivo Imaging System (PerkinElmer, Waltham, MA). Mice were also given weekly vaginal lavages using 200 μL of sterile PBS throughout the study to collect cells from the local microenvironment of the reproductive organs. Cells sourced from lavages were counted and spun down to remove PBS as it has been shown to suppress MALDI-TOF ionization.21 (link) This initial centrifugation step also removes mucous and mucous-associated proteins, such as mucin. Cells are then normalized to 10,000 cells/μL using DI water and stored at −80°C following collection. Upon addition of DI water, cells undergo osmotic stress and lyse, which removes the need for a wash step as all remaining proteins will be in solution. This lysing step prior to analysis also allows for proteins or other similarly sized molecules to be detected via MALDI-TOF MS. After two months of tumor progression, all animals were humanely sacrificed followed by collection of tumors and reproductive organs.
Xenogen ivis spectrum in vivo imaging system
Manufactured by PerkinElmer
Sourced in United States
The Xenogen IVIS® Spectrum In Vivo Imaging System is a laboratory instrument designed for non-invasive, real-time imaging of biological processes in living subjects. The system uses bioluminescence and fluorescence imaging techniques to detect and quantify biological signals within the animal models.
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10 protocols using xenogen ivis spectrum in vivo imaging system
1
Ovarian Cancer Tumor Progression Protocol
2
Breast Cancer Metastasis Imaging in Mice
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Luciferase expressing MDA-MB-231 cells were purchased from Perkin Elmer, Inc. NOD/Scid mice (001303; Jackson Laboratory, Inc.) or nu/nu mice (002019; Jackson Laboratory, Inc.) were used in this study. For the xenograft study of breast cancer metastasis, MDA-MB-231 breast cancer cells were exposed to mechanical strain or static conditions for 24 h. The cancer cells were either untreated or, treated with 10 μM of PI3-Kγ inhibitor (CAS 648450-29-7). We injected 5 × 105 cells were injected into the tail vein of 7-week-old female mice. The mice were given an intraperitoneal injection of luciferin (150 mg/kg) in DPBS 10 min prior to imaging. The luminescence was visualized using the Xenogen IVIS Spectrum In Vivo Imaging System (PerkinElmer).
3
Ovarian Tumor Growth Monitoring
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OVCAR8 RFP cells were either grown on traditional cell culture plastic or grown as MTS as described above for seven days. Cells (76,000) or MTS (38 MTS; 2000 cells per MTS) were collected, moved into phosphate buffered saline, and injected into the left ovarian bursa of 8-week old athymic nu/nu mice (Taconic Germantown, NY, USA). Tumor growth was imaged every two weeks starting at four weeks and every week starting at Week 12. Imaging was done with a Xenogen IVIS Spectrum In Vivo Imaging System (PerkinElmer, Waltham, MA, USA) as previously described [47 (link),48 (link)].
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Verticillin A Nanoparticle Therapy for Ovarian Xenografts
5
Monitoring Xenograft Tumor Growth
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Intravital Imaging of Joint Inflammation
7
Evaluating Anti-Tumor Effects of Taxol
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HeLa cells were labelled with luciferase (Shanghai Genechem Co.) using a lentiviral transduction method. luciferase-labelled HeLa cells were stably transfected with lentivirus carrying LIV-1 shRNA (HeLa /LIV-1 shRNA cells) or mock shRNA (HeLa/Mock cells) (Shanghai Genechem Co.). 1 × 106 tumour cells were implanted subcutaneously into the right flanks of Nod/SCID mice. The mice were injected i.p once a week (for four weeks) with Taxol (12 mg/kg) [27 (link),28 (link)] or DMSO when tumour sizes reached 200 mm3. The bioluminescence signal intensity (BLI) and tumour volumes were measured using the Xenogen IVIS spectrum in vivo imaging system (Caliper Life Sciences, Inc.). All animals were obtained from BEIJING HFK BIOSCIENCE Co., Ltd. (Beijing, China), and experiments were approved by the Committee on the Ethics of Animal Experiments of Tongji Medical College. Mice were maintained in the accredited animal facility of Tongji Medical College.
8
NF-κB Activation Monitoring in Mice
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All animal studies reported in this manuscript were performed in compliance with the UMN Institutional Animal Care and Use Committee under an approved protocol. The NGL or HLL mice have been used by our lab to describe the in vivo activation of NF-κB in various inflammatory states [37] (link), [38] (link). HLL mice were used to study the effects of intratracheal agmatine administration. Each mouse received an intratracheal injection of 100 µg agmatine in 100 µL of PBS via direct laryngoscopy while under isoflurane anesthesia. NF-κB activation at the timepoints indicated was performed as described with quantitation of luminescence over the chest using the Xenogen IVIS Spectrum in vivo imaging system (Caliper Life Sciences, Hopkinton, MA) for a 5 sec capture time. Each reading was normalized to the baseline luminescence of the same mouse before agmatine injection. The studies combining LPS and agmatine were performed in NGL mice as this line replaced the HLL mice in our colony during these studies. Agmatine and/or LPS were injected intraperitoneally (1 mg and 100 µg respectively in 200 µL PBS) and luminescence over the chest and abdomen was quantified and normalized on a per mouse basis.
9
NDV-Luci Infection in Mice
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Mice were infected with 5 × 107 PFU rNDV-Luci by tail intravenous injection every two days for three times. At 24 h post-final injection, mice were anesthetized with isoflurane mixed with oxygen and injected intraperitoneally with D-luciferin potassium salt (1.5 mg/mouse) for 10 min before imaging. Bioluminescence images were captured using Xenogen IVIS spectrum in vivo imaging system (Caliper Life Sciences, Hopkinton, MA).
10
Intramuscular mRNA Delivery and Bioluminescence Imaging
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