Ek. Hill et Aj. De Mello, Single-molecule detection using confocal fluorescence detection: Assessment of optical probe volumes, ANALYST, 125(6), 2000, pp. 1033-1036
We have constructed a simple component-based confocal detection system, tha
t is capable of fluorescence detection at the single-molecule level. The co
mponent-based format maximises flexibility and reduces start-up costs. A ne
w model for the optical probe volume is proposed, that is based on the 1/e(
2) Gaussian intensity contour of a laser beam focused to the diffraction li
mit. Observation of the onset of single-molecule detection in our experimen
tal system confirms that this model is more appropriate than a simple cylin
drical approximation for a probe volume defined by a wide, tightly focused
laser beam.