@article {2085, title = {Single molecules observed by near-field scanning optical microscopy. (With commentary)}, journal = {Science}, volume = {262}, year = {1993}, month = {1993 Nov 26}, pages = {1422-5}, abstract = {

Individual carbocyanine dye molecules in a sub-monolayer spread have been imaged with near-field scanning optical microscopy. Molecules can be repeatedly detected and spatially localized (to approximately lambda/50 where lambda is the wavelength of light) with a sensitivity of at least 0.005 molecules/(Hz)(1/2) and the orientation of each molecular dipole can be determined. This information is exploited to map the electric field distribution in the near-field aperture with molecular spatial resolution.

Commentary: A paper of many firsts: the first single molecule microscopy; the first extended observations of single molecules under ambient conditions; the first localization of single molecules to near-molecular precision (\ 15 nm), the first determination of the dipole axes of single fluorescent molecules; and the first near-molecular resolution optical microscopy, when a single fluorescent molecule was used to map the evanescent electric field components in the vicinity of a 100 nm diameter near-field aperture. Although eventually supplanted by simpler far-field methods, this paper ushered in the era of single molecule imaging and biophysics, and inspired the concept that would eventually lead to PALM. Even today, near-field single molecule detection lives on in the {\textquotedblleft}zero mode waveguide{\textquotedblright} sequencing approach promoted by Pacific Biosciences.

}, issn = {0036-8075}, doi = {10.1126/science.262.5138.1422}, url = {http://www.ncbi.nlm.nih.gov/pubmed/17736823}, author = {Betzig, E and Chichester, R J} }