Published: Vol 6, Iss 6, Mar 20, 2016 DOI: 10.21769/BioProtoc.1762 Views: 34796
Reviewed by: Vivien Jane Coulson-ThomasAnonymous reviewer(s)
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Abstract
Over the past decade, in vivo bioluminescent imaging has emerged as a non-invasive and sensitive tool for studying ongoing biological processes within living organisms (Contag et al., 1997; Contag et al., 1998). Based on the detection and quantitation of the photons produced by the oxidation of luciferin by luciferase enzymes (Harvey, 1927), this technique has proved to be particularly useful in analyzing cancerous cells and monitoring tumor growth (Edinger et al., 1999; Sweeney et al., 1999; Vidal et al., 2015), providing a cost-effective insight into how the disease progresses in vivo, without the need of serial sacrifice of animals. This protocol describes in detail the procedure of obtaining luciferase-tagged tumors in immunocompromised mice that can be studied by bioluminescent imaging through the use of an IVIS Spectrum imager.
Keywords: Bioluminescent imagingMaterials and Reagents
Equipment
Software
Procedure
Representative data
Figure 3. Luciferase detection in cells. Multiple methods can be used for testing the success of cell infection, which include but are not limited to: A. Genetic detection of luciferase gene by PCR and/or B. Testing luciferase activity.
Figure 4. Subcutaneous injection of cells in mice. A. Lift the skin over the back of the neck to make a tent. B. Insert the needle at the tent base, holding it parallel to the animal’s body to avoid puncturing underlying structures. C. Aspirate to create a light vacuum and ensure that the needle has not entered a blood vessel. Slowly inject the cell and extracellular matrix suspension.
Figure 5. Bioluminescence in vivo imaging. A. Representative luciferase time course (every 2 min) imaging of mice bearing subcutaneous tumors injected with luciferin. B. Graph illustrates the bioluminescence saturation of the whole mouse (displayed in A.) obtained until time point 8 min, when the experiment was decided to be concluded.
Acknowledgments
We thank the TJ Martell Foundation and Agilent Technologies for their support.
References
Article Information
Copyright
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Carceles-Cordon, M., Rodriguez-Fernandez, I., Rodriguez-Bravo, V., Cordon-Cardo, C. and Domingo-Domenech, J. (2016). In vivo Bioluminescence Imaging of Luciferase-labeled Cancer Cells. Bio-protocol 6(6): e1762. DOI: 10.21769/BioProtoc.1762.
Category
Cancer Biology > General technique > Tumor formation
Cancer Biology > Tumor immunology > Tumor formation
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