Mice were anaesthetised with isoflurane with 100 % oxygen and given an intraperitoneal injection of 15 mg/mL of d -luciferin (Gold Biotechnologies, ST Louis, USA). Mice were imaged after 5 min using a cooled charged-coupled device camera, (IVIS machine, Perkin Elmer, Coventry, UK) for between 1 s and 5 min. The regions of interest (ROI) were measured using Living Image Software (Perkin Elmer, Beaconsfield) and expressed as photons per second per centimetre squared per steradian (photons/second/cm2/sr).
Ivis machine
Manufactured by PerkinElmer
Sourced in United Kingdom
The IVIS machine is a laboratory imaging system designed for in vivo bioluminescence and fluorescence imaging. It allows researchers to visualize and quantify biological processes in living animals non-invasively. The IVIS machine captures high-sensitivity images and data that can be used to study a variety of applications, such as gene expression, cell trafficking, and disease progression.
Automatically generated - may contain errors
Lab products found in correlation
Living image software, by PerkinElmer (6 mentions)
Transit qr hydrodynamic delivery solution, by Mirus Bio (1 mentions)
Endo free maxiprep, by Qiagen (1 mentions)
Ivis imaging system, by Bruker (1 mentions)
Living image software, by Bruker (1 mentions)
D luciferin potassium salt solution, by GoldBio (1 mentions)
Living image software version 4, by PerkinElmer (1 mentions)
Isoflurane, by Abbott (1 mentions)
12 protocols using ivis machine
1
In Vivo Bioluminescence Imaging in Mice
2
In Vivo Bioluminescence Imaging
3
Longitudinal In Vivo Tumor Imaging
4
In Vivo Bioluminescence Imaging of Mice
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Where appropriate, mice were anaesthetised with isoflurane with 100% oxygen and/or received an intraperitoneal injection of 15 mg/ml of D-luciferin (Gold Biotechnologies, St. Louis, USA). Mice were imaged after 5 min using a cooled charged-coupled device camera (IVIS machine, Perkin Elmer, Coventry, UK) between 1 s and 5 min. The regions of interest were measured using the Living Image Software (Perkin Elmer) and expressed as photons per second per centimetre squared per steradian (photons/second/cm2/sr).
5
Bioluminescent Imaging in Mice
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At the indicated time post injection, mice were given 200 μl luciferin (15 mg/ml) intraperitoneally. Signal was allowed to stabilize for 10 min and then loaded into the Perkin Elmer IVIS machine for capture of luminescent signal (1 min exposure).
6
Luciferase Imaging of Hydrodynamic Gene Delivery
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Plasmid DNA was prepared by endo-free maxiprep (Qiagen). For long-term luciferase imaging, female FVB 8-week-old mice obtained from Baylor College of Medicine (Houston, TX) received hydrodynamic tail vein injections of TransIT-QR hydrodynamic delivery solution (Mirus Bio, Madison, WI) containing 25 μg of transposon DNA with or without transposase DNA (2.5 μg for pCMV-TcBuster or 5 μg for pCMV-piggyBac). Mice were imaged periodically under isoflurane on an IVIS machine (Perkin Elmer, Waltham, MA) as previously described (25 (link)). Mice that were sacrificed at 24 h were 4-month-old male FVB mice obtained from Charles River. Hydrodynamic tail vein injections contained 10 μg of pTcBCAGLuc and either 2.5 μg or 25 μg of pCMV-HA-TcBuster (n = 3 per group) in 2 ml of TransIT-QR solution. Luciferase positive mice (n = 2 per group) were sacrificed 24 h post-injection and their livers fixed in 10% formalin at 4°C overnight. Samples were stained for the HA antigen as previously described (29 (link)). The Institutional Care and Use Committees of Baylor College of Medicine and Vanderbilt University approved all animal experiments. Error bars represent the standard error of the mean.
7
Liver-targeted CRISPR-Cas9 Tumor Induction
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Liver-specific AAV-CRISPR vectors were designed to co-cistronically express FLuc and Cre recombinase for induction of Cas9 expression under a TBG promoter when delivered to LSL-Cas9 mice (plasmids available at Addgene). Two sgRNA expression cassettes were built in these vectors, one encoding an sgRNA targeting Trp53, with the other being an open sgRNA cassette (double Sap I sites for GTS cloning). The vector was generated by gBlocks Gene Fragment synthesis (IDT) followed by Gibson assembly (NEB). Each specific sgRNA targeting a driver gene was cloned separately into this vector. AAV9 virus was produced and qPCR-titrated, as described above. Total viral particles (1 × 1011) were introduced by intravenous injection into LSL-Cas9 mice. For combinations of two AAVs, 5 × 1010 viral particles were used from each AAV to generate equal titer mixtures and injected. Four to six mice were injected per group. Starting 1 month after injection, mice were imaged by IVIS each month. Briefly, mice were anesthetized by intraperitoneal injection of ketamine (100 mg/kg) and xylazine (10 mg/kg) and imaged for in vivo tumor growth using an IVIS machine (PerkinElmer) with firefly d -luciferin potassium salt (150 mg/kg body weight) injected intraperitoneally. Relative luciferase activity was quantified using Living Image software (PerkinElmer).
8
In Vivo Bioluminescence Imaging
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Mice were anesthetized by isoflurane inhalation and imaged for metastasis using an IVIS machine (PerkinElmer) 5 minutes following intraperitoneal injection of firefly d-luciferin potassium salt (150 mg/kg body weight).
9
Murine Tumor Models for Immunotherapy
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All experiments were performed according to the institutional guidelines for the care and use of animals and were approved by the Animal Care and Use Committee of the Second Xaingya Hospital, Central South University. To establish LLC mouse models with subcutaneous tumors and metastatic tumors, LLC-sgNTC and LLC-sgAtrx cells were suspended in 150 μl of PBS at a density of 1 × 106/ml. Then, these cell suspensions were subcutaneously injected or intravenously injected via the tail vein into C57BL/6 mice with a 1-ml syringe. The anti-CTLA4 and anti–PD-1 were given at a dose of 200 ug/mice at 9, 12, and 15 days after the establishment of models. The sizes of the subcutaneous tumors were measured by Vernier calipers every 3 days [tumor volume = 1/2 × (L × W)2]. For the metastasis model, tumor volume was monitored in vivo by bioluminescence detected by the IVIS imaging system (Bruker, USA) once every month. A D-luciferin potassium salt solution (Goldbio. St. Louis, MO, USA) was injected intraperitoneally (150 mg/kg), and 10–15 min after injection, the mice were imaged for in vivo tumor growth using an IVIS machine (PerkinElmer). Living Image Software (Bruker MI, USA) was used to measure the total flux of the metastatic lung tumor.
10
In Vivo Bioluminescence Imaging of Mice
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Where appropriate, mice were anaesthetised with isoflurane with either 100% oxygen or medical air (21% oxygen) (Abbotts Laboratories, London, UK) and/or received an intraperitoneal injection of 15 mg/mL of D-luciferin (Gold Biotechnologies, ST Louis, USA). Mice were imaged after 5 minutes using a cooled charged-coupled device camera, (IVIS machine, Perkin Elmer, Coventry, UK) for between 1 second and 5 minutes. The regions of interest (ROI) were measured using Living Image Software (Perkin Elmer) and expressed as photons per second per centimetre squared per steradian (photons/second/cm2/sr).
The imaging chambers used to image conscious mice for the intracranial injected mice had the following dimensions; 5 cm × 5 cm × 6 cm. The Perspex box which was used to image the conscious mice which had received an intravenous injection is shown in Fig.2 .
The imaging chambers used to image conscious mice for the intracranial injected mice had the following dimensions; 5 cm × 5 cm × 6 cm. The Perspex box which was used to image the conscious mice which had received an intravenous injection is shown in Fig.
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