Nucleofector 96-well Shuttle From Amaxa Biosystems

The Nucleofector is essentially an electroporator that permits efficient transfection of primary cells and cell lines. This non-viral method relies on a device, the Nucleofector device, which electroporates cells while in a proprietary, cell-type-specific Nucleofector solution. The Nucleofector 96-well Shuttle allows transfection in a 96 wells format; the Shuttle is simply an extension to the Nucleofector II device which is connected to a laptop computer. In addition to the obvious advantage for high-throughput screening, the 96-well format allows the experimenter to use a substantially lower number of cells per condition (about 1:5). This fact can be critical when working with hard-to-harvest primary cell lines.

The 96-well shuttle uses very accessible software that nonetheless lacks some basic OS functions. For instance, although each well can be transfected using a different program, the CTRL key does not allow you to select staggered wells. Besides some other inconsequential annoyances, both software and hardware are reliable and user-friendly.

In my opinion, the 96-well shuttle suffers from three drawbacks. First, the instrument utilizes proprietary electrical parameters to electroporated the cells. Hundreds of program combinations exist, and one is certainly optimal to transfect your favorite cell type. Unfortunately, troubleshooting is limited, because the electrical parameters, despite their likely simplicity, are undisclosed by the manufacturer. You will have to contact the company technical support (which is actually helpful and knowledgeable) who will recommend programs to use in preliminary optimization experiments. Second, the plasmid preparation used for electroporation needs to be of excellent quality: endotoxin-free preparations of ~0.5 mg/ml are recommended. Last, the product can be considerably more costly than alternative transfection techniques.

With these concerns in mind, I admit that I have been confidently using the Amaxa technology for the last 4 years, and unreservedly switched to the 96-well shuttle in the last 6 months. Several attributes make nucleofection my preferred method of protein expression: (1) transfection efficiency and level of protein expression is extremely high. I consistently obtain 30-50% transfection efficiency in primary neuronal cultures, (2) low toxicity compared to viral-based infection, (3) protein expression starts as early as six hours post-nucleofection, (3) in my experience, electroporation is particular useful when trying to transfect multiple plasmids into single cells. When performing triple transfection, I find that >50% of the cells are triple transfected, as opposed to <20% when using lipid-based reagents, (4) the transfection efficiency reported on their website actually matches results that you will obtain (typically 40-80% efficiency for hard to transfect cells), (5) reproducibility from experiment to experiment is high, (6) when used to transfect siRNA, transfection efficiency reaches 99%. One tip: when transfecting neurons, I suggest using 3-10 fold the number of cells recommended in their protocol, particularly for cortical neurons. Perhaps I should work as an Amaxa sales representative!