![]() This is achieved by a similar arrangement: the emission light is fed through a tiny aperture, the detection pinhole, before recorded by a sensing element. That is, the sensing region should as well be a diffraction limited spot, at all times coinciding with the illumination spot. On the detection side, the sensor must follow in an equal manner. As all conventional light sources are usually not spot-shaped but have a significant extension, the light source is projected on a tiny aperture, the pinhole, acting as a spot-shaped source. The psf is the distribution of light in the focus of an optical device when imaging a dimensionless spot. A diffraction limited spot in the focal plane of the microscope is the image of a spot-shaped light source and therefore called “point spread function, psf”. The smallest shape that can be generated is a diffraction limited spot. To achieve this task, the sample is scanned by a tiny light point. Optimally, the slices cover just the depth of focus generated in the microscope. Confocal microscopes allow to optically separate slices of the sample’s response and record theses slices as images.
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