Point Scanning Microscope with Adaptive Illumination Beam Intensity

Abstract

In this paper we describe a point scanning optical microscope where the illumination beam can be programmably controlled in real time using a liquid crystal spatial light modulator (LCSLM). With an appropriate pattern displayed on the LCSLM, the device can be made to act as a binary diffraction hologram. In the proposed microscope the illumination beam is in fact the +1 order beam diffracted from the binary hologram. By displaying a sequence of binary holograms it is possible to make a beam scanning, similar to a conventional scanning microscope. Here we use a computer generated holography technique to compute the binary holograms which facilitate complete control of the amplitude and phase profile of the illumination beam. In a number of microscopy applications using reflected light, the reflectivity of the sample plane may differ from region to region. Therefore if a single illumination beam intensity is used for the whole sample plane, then the regions with less reflectivity will be imaged with poor signal to noise ratio. In our proposed microscope the sample plane is first imaged to determine the regions of weak reflectivity. Holograms are then computed to make the illumination beam adapt to the reflectivity variations in the given sample plane. The image obtained with the modified set of holograms have superior signal to noise ratio all over, relative to a conventional point scanning microscope with a fixed intensity illumination beam. In this paper we present some preliminary results using the proposed setup.