In Vivo Bioluminescence Imaging of Cells

Bioluminescence imaging with an IVIS was conducted as described previously.⁵⁷ (link) In brief, eight mice were infected with 1 × 10¹⁰ CFU STEC E32511 or O111 (four mice for each strain). After 1 or 2 days, mice were intravenously injected with nano-lantern-labeled Muse or non-Muse cells (1 × 10⁵/mouse).²⁵ (link) Mice were then intravenously injected with 100 μL substrate (80 μg/mL coelenterazine-h; Wako, Tokyo, Japan), anesthetized with isoflurane by inhalation, and placed in an acrylic plastic box. Bioluminescent images were acquired using an IVIS (Spectrum; PerkinElmer, Waltham, MA, USA) using the following settings: exposure time, 1 min; medium binning, F/Stop = 1. Data acquisition and analysis were performed using Living Image 4.4 software (PerkinElmer). Quantification was performed using a region of interest (ROI) defined manually (abdominal center or injection site), and the results are expressed as photons per second (P/s).

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Ozuru R., Wakao S., Tsuji T., Ohara N., Matsuba T., Amuran M.Y., Isobe J., Iino M., Nishida N., Matsumoto S., Iwadate K., Konishi N., Yasuda K., Tashiro K., Hida M., Yadoiwa A., Kato S., Yamashita E., Matsumoto S., Kurozawa Y., Dezawa M, & Fujii J. (2019). Rescue from Stx2-Producing E. coli-Associated Encephalopathy by Intravenous Injection of Muse Cells in NOD-SCID Mice. Molecular Therapy, 28(1), 100-118.

Publication 2019

coelenterazine-h Living Image 4.4 software

Abdominal Cells Coelenterazine Inhalation Isoflurane Mice Muse Stec Strain

Corresponding Organization : Tottori University

Other organizations : Tohoku University, Okayama University, Hasanuddin University, Toyama Institute of Health, University of Toyama, Jikei University School of Medicine, Tokyo Metropolitan Institute of Public Health, Kyushu University, Tottori University Hospital, Niigata University

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Variable analysis

independent variables

Mouse strain (STEC E32511 or O111)

dependent variables

Bioluminescent signal (photons per second, P/s)

control variables

Number of mice (4 per strain)
Dose of bacteria (1 × 10^10 CFU)
Time of cell injection (1 or 2 days after bacterial infection)
Cell type (nano-lantern-labeled Muse or non-Muse cells, 1 × 10^5/mouse)
Substrate concentration (100 μL of 80 μg/mL coelenterazine-h)
Imaging settings (exposure time, binning, F/Stop)
Anesthesia (isoflurane)

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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