LUC;CCSP.CRE mice, bioluminescent reporters of CCSP-labeled cell mass, received one intraperitoneal injection of saline (100 μL saline) or urethane (1 g/Kg in 100 μL saline) and were serially imaged before treatment start, and at 150 and 210 days into treatment. Imaging was done on a Xenogen Lumina II (Perkin-Elmer, Waltham, MA) 5–20 min after delivery of 1 mg D-Luciferin sodium in 100 μL of sterile water to the retro-orbital vein, and data were analyzed using Living Image v.4.2 (Perkin-Elmer, Waltham, MA) (Stathopoulos et al., 2007 (link); Vreka et al., 2018 (link)).
Xenogen lumina 2
Manufactured by PerkinElmer
Sourced in United States
The Xenogen Lumina II is a laboratory imaging system designed for in vivo bioluminescence and fluorescence imaging. It provides a versatile and sensitive platform for a range of applications, such as small animal imaging and cell-based assays.
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4 protocols using xenogen lumina 2
1
Bioluminescent Imaging of CCSP-Expressing Cells
2
Bioluminescent Imaging of Cells and Mice
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Living cells and mice were imaged 0, 4, 8, 24, and 48 h after cellular treatments and 0 h, 4 h, and 12–14 days after pleural delivery of pNGL-expressing cells on a Xenogen Lumina II (Perkin-Elmer, Waltham, MA) after addition of 300 μg/mL D-luciferin to culture media or isoflurane anesthesia and delivery of 1 mg intravenous D-luciferin to the retro-orbital veins8 (link)–11 (link),16 (link),25 (link),34 (link),35 (link). Data were analyzed using Living Image v.4.2 (Perkin-Elmer).
3
In Vivo and In Vitro NF-κB Bioluminescence
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Mice were imaged for NF-κB reporter bioluminescent signal daily starting at day 10 post-tumor cell injection until sacrifice. For this, mice were anesthetized by isoflurane inhalation and were imaged for bioluminescence on a Xenogen Lumina II (PerkinElmer, Waltham, MA, USA) 5–20 min after delivery of 1 mg D-Luciferin potassium salt diluted in 100 μL of sterile water into a retro-orbital vein. Pleural tumors isolated from NGL mice were also imaged ex vivo for green biofluorescence using 410–440 nm background control excitation, 445–490 nm experimental excitation, and 515–575 nm emission passbands on a Xenogen Lumina II. Cells were imaged for bioluminescence on a Xenogen Lumina II 0, 4, 8, 16, and 24 h after a single addition of 300 μg/mL (equivalent to 1 mM) D-luciferin to the culture media. Data were analyzed using Living Image v.4.2 (PerkinElmer, Waltham, MA, USA) as described previously [10 (link),11 (link),20 (link),21 (link),22 (link),34 (link)].
4
In vivo Cre-recombinase Adenoviral Delivery
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Isoflurane‐anesthetized C57BL/6 and mT/mG mice received 5 × 108 PFU intrapleural or intraperitoneal Ad‐Cre or Ad‐Luc in 100 μl PBS and were serially imaged for bioluminescence on a Xenogen Lumina II (Perkin‐Elmer, Waltham, MA) after receiving 1 mg retro‐orbital D‐luciferin under isoflurane anesthesia, and data were analyzed using Living Image v.4.2 (Perkin‐Elmer; Stathopoulos et al, 2006 (link); Spella et al, 2019 (link)), or were euthanized and pleural lavage was performed, lungs were explanted, and parietal pleura was stripped. For pleural lavage, 1 ml PBS was injected, was withdrawn after 30 s, and was cytocentrifuged onto glass slides (5 × 104 cells, 300 g, 10 min) using CellSpin (Tharmac, Marburg, Germany). Lungs were embedded in optimal cutting temperature (OCT; Sakura, Tokyo, Japan) and sectioned into 10‐µm cryosections. The parietal pleura was placed apical side up onto glass slides. Samples were stained with Hoechst 55238 and were examined on AxioObserver D1 (Zeiss, Jena, Germany) or TCS SP5 (Leica, Heidelberg, Germany) microscopes.
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