The iBox® Scientia animal imaging system available from Analytik-Jena can be used to perform in vivo fluorescence imaging of proteins. The imaging system has been developed for imaging a broad range of animals. Multiple label studies can be conducted by using the extensive range of fluorescent proteins that have different emission spectra.
The proteins used with the system are intrinsically fluorescent, avoiding the need for cofactors and substrates. Moreover, Analytik-Jena has been constantly optimizing the fluorescent behavior of reporter proteins, with a wide array of bright red fluorescent proteins available recently. The genes for these proteins can be purchased by academic and commercial institutions.
Expressed Fluorescent Proteins
Together with Analytik-Jena’s f1.2 optics, the iBox® Scientia can be used for fast exposures, thus enabling the capture of precise images that remain unaffected by the animal’s breathing. The VisionWorks software from Analytik-Jena’s can be used with potently fluorescent proteins to identify the margins of tumors and to measure the size of the tumor.
In a study performed by Zhao et al., a Salmonella typhimurium mutant was injected every day into specimen. S. typhimurium builds up in tumors and minimizes (or totally prevents) the metastatis of PC-3 prostate tumor cells in humans. In that study, human glioma cells were expressed by using a red fluorescent protein (RFP), named U87.
Figure 1. iBox® Scientia small animal imaging system. Image Credit: Analytik-Jena US.
Materials and Methods
The iBox Scientia® in vivo imaging system from Analytik-Jena was configured with the features listed below:
- BioLite™ automated excitation light source
- Motorized sample platen
- f1.2 optics—large-area, fast biolens
- High-resolution (4.2 mpx) BioChemi 500 camera
- CFP, GFP, RFP, and YFP emission and excitation filter sets
- VisionWorks® LS analysis software
- Analytik-Jena Heater, set to a temperature of 370 °C
A simple imaging technique was employed. A small volume of U87 cells was injected into nude mice through subcutaneous injection. Day 0 was labeled at the point where the tumor was detected to have grown to a size of 1 mm2. An injection of 5 x 107/100 µL S. typhimurium A1-R was administered to the treatment group via the tail on a weekly basis.
Figure 2. RFP orthotopic tumors in nude mice imaged over a period of two weeks using Analytik-Jena iBox® Scientia small animal imaging system. Image Credit: Analytik-Jena US.
White light and fluorescence images were captured by using the excitation and emission filter for RFP together with the Analytik-Jena iBox®. Then, these images were combined through blend control (where the source and destination were set at 75% and 25%, respectively).
The fluorescent image was colored red to pinpoint the position at which RFP was present. The exposure times used for white light and RFP images were 0.01 seconds and 1 second, respectively.
Results
The untreated controls and the treated group were compared after a week and two weeks, revealing that there was a decrease in tumor growth in the treated group. This indicates that tumor growth can be reduced by administering weekly injections of S. typhimurium.
Conclusion
Studies of tumor imaging using fluorescent imaging, similar to the one described in this article, point toward the significance of an accurate process in time course research, as well as that of fluorescent protein imaging during in vivo analysis of cancers. The iBox® Scientia integrated with the f1.2 optics available from Analytik-Jena provides sharp images that remain unaffected by the breathing of the subject that is imaged.
References
- Hoffman, R.M. and Yang, M. Whole-body imaging with fluorescent proteins. Nat. Protoc. 2006, 1, 1429–1438.
- Hoffman, R.M. and Yang, M. Color-coded fluorescence imaging of tumor-host interactions. Nat. Protoc. 2006, 1, 928–935.
- Zhao, M.; Geller, J.; Ma, H.; Yang, M.; Penman, S.; Hoffman, R.M. Monotherapy with a tumor-targeting mutant of Salmonella typhimurium cures orthotopic metastatic mouse models of human prostate cancer. Proc. Natl. Acad. Sci. U. S. A. 2007, 104, 10170–10174.
- Zhao, M.; Yang, M.; Ma, H.; Li, X.; Tan, X.; Li, S.; Yang, Z.; Hoffman, R.M. Targeted therapy with a Salmonella typhimurium leucine-arginine auxotroph cures orthotopic human breast tumors in nude mice. Cancer Res. 2006, 66, 7647–7652.
About Analytik Jena US
Analytik Jena is a provider of instruments and products in the areas of analytical measuring technology and life science. Its portfolio includes the most modern analytical technology and complete systems for bioanalytical applications in the life science area.
Comprehensive laboratory software management and information systems (LIMS), service offerings, as well as device-specific consumables and disposables, such as reagents or plastic articles, complete the Group’s extensive range of products.
About Life Science
The Life Science product area demonstrates the biotechnological competence of Analytik Jena AG. We provide a wide product spectrum for automated total, as well as individual solutions for molecular diagnostics. Our products are focused to offer you a quality and the reproducibility of your laboratory results. This will surely ease your daily work and speed up your work processes in a certain way.
All together we support you through the complete process of the lab work. Besides we offer customized solutions and are able to adapt our products to your needs. Automated high-throughput screening systems for the pharmaceutical sector are also part of this segment’s extensive portfolio.
About Analytical Instrumentation
Analytik Jena has a long tradition in developing high-performance precision analytical systems which dates back to the inventions made by Ernst Abbe and Carl Zeiss. We have grown to become one of the most innovative manufacturers of analytical measuring technology worldwide.
Our business unit Analytical Instrumentation offers excellent competencies in the fields of optical spectroscopy, sum parameters and elemental analysis. Being proud of our core competency we grant all our customers a long-term warranty of 10 years for our high-performance optics.
About Lab Automation
With more than 25 years of market experience, Analytik Jena with its CyBio® Product Line is a leading provider for high quality liquid handling and automation technologies. In the pharmaceutical and life science industries, our products enjoy the highest reputation for precision, reliability, robustness and simplicity.
Moreover, the Automation Team designs, produces and installs fully automated systems tailored to our clients' application, throughput and capacity requirements. From stand-alone CyBio® Well up to fully customized robotic systems we handle your compounds, biomolecules and cells with great care.
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