Compensating the effects of light attenuation in confocal microscopy by histogram modelling techniques

Confocal Laser Scanning Microscopy (CLSM) enables us to capture images representing optical section on the volume of a specimen. The images captured from different layers have a different contrast: the images obtained from the deeper layers of the specimen will have a lower contrast and will present a certain degree of blur in respect to the images obtained from the topmost layers. The main reasons responsible for the effects described above are light absorption and scattering by the objects and particles that are present in the volume through which the light passes. Also light attenuation can be caused by the inclination of the observed surface. In the case of the surfaces that have a steep inclination the reflected light will have a different direction than the one of the detector. We propose a technique that can be used to compensate the effects of light attenuation based on histogram operations. We will consider two types of histogram operations: histogram equalization and histogram specification. A discussion of the results that were obtained using the two types of operations will be presented.