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OMICS Transfection Continued from page 33 or plasmid, purity of pDNA, and media. Nonchemically Mediated Methods The age-old transfection pitfalls remain. Cells die after they are transfected, or cells remain healthy only because they are not transfected. "Increasing the transfection effciency will simplify and clarify experimental answers. For example, if you have a population of cells that are only 50% affected, it can be diffcult to determine the true response to treatment," said Steve Kulisch, cell biology business unit manager at Bio-Rad. "Transfection effciency rates with many immortalized cell lines are quite good. However, researchers continue to adopt additional, more challenging cell lines in their work, forcing the scientifc-tools community to keep up." The desire to get more effcient delivery of biomolecules forces innovation. With the vast number of cells researchers use today, the Holy Grail is to fnd some technology or technique that is effcacious for all cell lines with minimal amounts of optimization. Cellular variations include: varying lipid compositions in the plasma membranes and different cell sizes with ranging amounts of When was the last time you looked at your cells? Don't just read. Visualize! Get rich cellular resolution assays and well-based intensity readings in one small footprint. The SpectraMax® MiniMax™ Imaging Cytometer option for the SpectraMax® i3 Multi-Mode Detection Platform is specifically designed to: • Inspect samples quickly with brightfield and fluorescent imaging • Lower variance by normalizing readings with number of cells per well • Satisfy plate reading and imaging workflow in one instrument Minimize workflow hassle. Maximize time and results. The SpectraMax i3 System (top, blue) with MiniMax™ Imaging Cytometer (bottom, orange) Together through life sciences. exposed surface areas. Adherent versus suspension cultures affect surface area as well. Getting the material into the cell is step one. Step two is getting that biomolecule imported to where it needs to be; some cells are more effcient at biomolecule transport than others. Bio-Rad pioneered electroporation as a transfection methodology and also offers biolistic devices. These devices coat small gold or tungsten microparticles with nucleic acids or proteins, then use a helium blast to accelerate the microparticles into the sample. Although the company's technologies have not changed dramatically since their introduction, a recently introduced electroporation buffer assists with opening up the pores on the cells and also provides energy to help with recovery, improving electroporation transfection effciency. Many basic research questions are still frst addressed by transfecting easy-to-access and easy-to-culture, but also artifcially immortalized or cancer-derived, cell lines. To confrm that insights gained from these "artifcial" cell line systems refect the in vivo situation, there is a need to switch to more biologically relevant models. For therapeutic approaches, the use of primary animal cells could reduce the number of animal tests, and use of primary human cells allows extrapolating fndings from such animal models. "The challenge for transfection of biologically relevant cells, such as primary and stem cells, is to fnd an easy-to-use method that achieves reproducible high transfection effciencies without major impacts on viability and functionality," explained Andrea Toell, Ph.D., senior product manager at Lonza Pharma & Biotech-Bioscience Solutions. A technique that holds promise, viral transduction typically achieves high effciencies in primary cells and allows for stable integration. But working with viruses is laborious. Some viruses may also have an infuence on functionality due to their stable integration and induction of interferon responses. Lonza's electroporation-based Nucleofector Technology is a non-viral technology It achieves transfection effciencies close to virus transduction for hard-to-transfect cells, according to a company offcial, who added that compared to classical electroporation, the technology achieves much higher viabilities for primary cells and higher effciencies with smaller amounts of DNA. Transfection reagents require cell proliferation for transferring the DNA into the nucleus for expression, while the Lonza technology allows transfection of nondividing cells, such as unstimulated T cells or neurons, as the DNA is directly transferred to the nucleus. Large-Scale Transfections Scan the QR code to learn more or visit www.moleculardevices.com/visualize See us at MipTec 2013, Booth #D33 For research use only. Not for use in diagnostic procedures. ©2013 Molecular Devices, LLC. The trademarks mentioned herein are the property of Molecular Devices, LLC or their respective owners. Another key aspect of transient transfection innovation is the ability to simply and cost-effectively scale transfections without sacrifcing performance or primary cell compatibility. See Transfection on page 36 34 | September 1, 2013 | GENengnews.com | Genetic Engineering & Biotechnology News