A research team comprised of Alexandro Pertsinidis, Yunxiang Zhang and Steven Chu, a Nobel laureate and US Energy Secretary, has developed an optical microscopy technique for making nanometre-scale measurements.
The researchers used two back-illuminated Andor EMCCD cameras: the iXon+ 860, capable of acquiring images at 500fps, and the highly sensitive iXon+ 897 model, which can capture individual photons from single fluorescence emitters.
By developing an active feedback system, the team were able to repeatedly place the centroid of a single fluorescent molecule image anywhere on either of the camera's CCD arrays and measure its position with sub-pixel accuracy. This means, that in conjunction with additional optical beams to stabilise the microscopy system, the traditional errors caused by non-uniformity of chip manufacture can be reduced to sub-nanometre scale.
As a result, the team were able to develop a two colour, single molecule imaging system, which achieved image resolutions with an order of magnitude greater than the current best super resolution techniques (5nm).
Gaining the ability to resolve single molecules at this level has significant implications for biological research, where it should allow the structure of large, multi-subunit complexes to be analysed at the single molecule level.
Chu and his colleagues are planning to harness the new technique to learn more about the human RNA polymerase II system, which initiates the transcription of DNA, and the molecular mechanisms controlling cell-to-cell adhesion processes.
The resolving power of this super resolution technique may also be of use in guiding the design of new photometric imaging systems in scientific fields such as nanometrology, atomic physics and astronomy.
