A team of researchers at Boston University (Boston, MA, USA) has, for the first time, been able to demonstrate the feasibility of high-speed optical identification of individual converted DNA bases as they translocate through solid state nanopores with high temporal resolution (1000 frames/s).
The team, which is led by Amit Meller, associate professor of biomedical engineering and physics, based their work on a custom total internal reflection fluorescence (TIRF) microscopy set-up, incorporating an ultra-sensitive Andor iXon 860 EMCCD camera to rapidly record fluorescence images from the nanopore membrane.
Meller believes the applicability of this technique has major implications for future approaches to DNA sequencing, “At between 50-250 bases per second, our DNA readout speed is already faster than other single molecule methods, but we believe we can push this up to greater than 500 bases per second by adapting the technique for 4-colour analysis and optimizing the reagents. After that, since our readout process does not involve an enzyme, the speed per nanopore will be determined by the limits of detection offered by state-of-the-art CCD or CMOS technologies, and is readily controlled by the voltage we apply on the SiN membrane. As soon as progress is made in raising imaging speeds, we can immediately utilize in our system, further increasing the readout rates.”
He adds that, “The use of highly sensitive and ultra-fast EMCCD is central to our sequencing method, as we rely on fast multi-colour optical readout, from many nanopores simultaneously."
For more information on the research, click HERE.
Posted by Vision Systems Design