We confirmed luciferase expression and activity in transduced cells with in vitro bioluminescence imaging. 10,000, 15,000, and 20,000 GBM cells/well were plated in black clear-bottom 96-well plates (Corning, Corning, NY, USA). The medium was replaced with 100 µl DPBS supplemented with 5 mM D-glucose and 1 mM sodium pyruvate (MilliporeSigma, St. Louis, MO) after 24 h. XenoLight® RediJect™ D-Luciferin (150 µg/ml; PerkinElmer, Waltham, MA, USA) was diluted 1:100 in DPBS supplemented with 5 mM D-glucose and 1 mM sodium pyruvate. Immediately after adding 100 µl of XenoLight® RediJect™ D-Luciferin dilution to each well (1:100 dilution, 0.15 µg/well), bioluminescence images were acquired with an IVIS® Spectrum (FOV: 13 cm, f-stop: 1, binning: 4; 124,262, PerkinElmer, Waltham, MA, USA). Bioluminescence was recorded every minute for 5 min, then every 5 min for 25 min. Bioluminescence was analyzed with Living Image 4.7.3 (PerkinElmer, Waltham, MA, USA) using a 96-well grid ROI. Bioluminescence for each well was expressed as total photons/s.
Xenolight rediject d luciferin
Manufactured by PerkinElmer
Sourced in United States
XenoLight® RediJect™ D-Luciferin is a bioluminescent substrate used for in vivo imaging of luciferase-expressing cells or organisms. It is a purified form of the firefly luciferase substrate, D-luciferin.
Automatically generated - may contain errors
Lab products found in correlation
Living image 4, by PerkinElmer (2 mentions)
Imagem2, by Hamamatsu Photonics (1 mentions)
Fusion fx7 edge, by Vilber (1 mentions)
Xenogen ivis, by PerkinElmer (1 mentions)
Ivis spectrum ct instrument, by PerkinElmer (1 mentions)
Living image software, by PerkinElmer (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Ivis spectrum, by PerkinElmer (1 mentions)
6 protocols using xenolight rediject d luciferin
1
Bioluminescent Imaging of Transduced Cells
2
Lumbar Spinal Cord Luminescence Imaging
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150 μL of XenoLight RediJect D-luciferin (30 mg mL− 1; PerkinElmer, Waltham, MA, USA) was injected intraperitoneally into HCN4Luc/tTA_TRE mice. The mouse was deeply anesthetized 30 min after the injection, and the lumbar spinal cord was dissected out. Transverse sections (400 μm thickness) of the spinal cord were obtained using a microslicer (Linear Slicer PRO 1; Dosaka EM, Kyoto, Japan). Luminescent imaging was performed using an EM-CCD camera (ImagEM2; Hamamatsu Photonics, Hamamatsu, Japan).
3
Therapeutic Efficacy of SHED-TK in Glioblastoma
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Forty female nude mice were anesthetized, and the tumor and SHED-TK (H1299 + SHED-TK) or tumor cells (H1299 only) were implanted as described above. In the coimplantation group, 5 × 104 H1299-RFP-luc2 cells mixed with the same number of SHED-TK cells in 5 μL RPMI 1640 were implanted into the right brain of nude mice. In the tumor implantation group, 5 × 104 H1299-RFP-luc2 cells in 5 μL RPMI 1640 were similarly implanted. The mice were intraperitoneally injected with GCV (50 mg/kg body weight in 200 μL PBS) or PBS twice per day from day 1 after tumor implantation for 10 consecutive days.9 (link)
Biological luminescence imaging of H1299-RFP-luc2 was performed by Fusion FX.7 EDGE (Vilber-Lourmat) on days 1, 15, and 29 after tumor implantation. The luminescence signals were measured by a charged-coupled device (CCD) camera in Fusion FX.7 10 min after the intraperitoneal injection of 150 mg/kg XenoLight Rediject D-luciferin (PerkinElmer, Waltham, MA). The exposure time was 2 min.
Biological luminescence imaging of H1299-RFP-luc2 was performed by Fusion FX.7 EDGE (Vilber-Lourmat) on days 1, 15, and 29 after tumor implantation. The luminescence signals were measured by a charged-coupled device (CCD) camera in Fusion FX.7 10 min after the intraperitoneal injection of 150 mg/kg XenoLight Rediject D-luciferin (PerkinElmer, Waltham, MA). The exposure time was 2 min.
4
Bioluminescent Lung Cancer Model
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Lung cancer models of NSCLC were created by injecting 5 × 106 A549-luciferase cells in sterile PBS into the tail veins of female SCID beige mice. Cancer development and progression were monitored using the Xenogen IVIS (PerkinElmer) bioluminescence imaging system. 100 μL of 30 mg/mL Xenolight Rediject D-Luciferin (PerkinElmer) was injected intraperitoneally into the mice approximately 10 min before the animals were imaged.
5
Luciferase Expression Monitoring in Mice
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Mice that were administered a total of 1 × 1011 vector genomes of AAV6.2FF-Luc or AAV6.2FF-hACE2 intranasally (Section 2.3 ) were used for detection of luciferase expression. Bioluminescence imaging was performed on days 3, 5, 7, and 10 after intraperitoneal (IP) administration of XenoLight RediJect D-Luciferin (Perkin Elmer, Waltham, MA, USA) at a concentration of 150 mg per kg using the IVIS SpectrumCT instrument (Perkin Elmer, Waltham, MA, USA). Resultant data were analysed and the signal intensity quantified using Living Image software (Perkin Elmer, Waltham, MA, USA).
6
Bioluminescent Imaging of Tumor Growth
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Bioluminescent imaging was initiated 7 days after implantation of Raji-Luc/GFP tumor cells and conducted weekly thereafter. Each mouse received an intraperitoneal injection of 100 μl (3 mg) of Xenolight Rediject d -luciferin (PerkinElmer, Waltham, MA) and maintained under isoflurane gas anesthesia for 5 min before imaging. Bioluminescent images were obtained using the Xenogen IVIS Spectrum (Caliper Life Sciences, Hopkinton, MA). Quantification of tumor bioluminescent activity was performed with Living Image 4.4 software (PerkinElmer, Waltham, MA) by taking an auto-exposure and recording the total photons in a set region of interest that was kept consistent throughout the duration of study. Bioluminescent activity is presented as photons per second per cm2 per steradian (p/s/cm2/sr). Additional sets of images were taken at 60-s exposures for display purposes in their respective figures.
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