Image scanning microscopy with multiphoton excitation or Bessel beam illumination

Colin JR Sheppard, Marco Castello, Giorgio Tortarolo, Eli Slenders, Takahiro Deguchi, Sami V Koho, Giuseppe Vicidomini, Alberto Diaspro (see publication in Journal )

Abstract

In image scanning microscopy, the pinhole of a confocal microscope is replaced by a detector array. The point spread function for each detector element can be interpreted as the probability density function of the signal, the peak giving the most likely origin. This thus allows a form of maximum likelihood restoration, and compensation for aberrations, with similarities to adaptive optics. As an example of an aberration, we investigate theoretically and experimentally illumination with a vortex doughnut beam. After reassignment and summation over the detector array, the point spread function is compact, and the resolution and signal level higher than in a conventional microscope.