Image contrast restoration in total internal reflection fluorescence microscopy

Blurring is a fundamental constraint in images of an optical imaging system. It appears because of interactions of light with the applied optical system and therefore, deblurring depends on features of the optical imaging system. We applied the iterative restoration method of Richardson–Lucy on the image that was taken with a total-internal-reflection-fluorescence microscope and found it is a very efficient method in improving the contrast of images if, the signal level of fluorophore emission in an image is considered and in an area with a low signal level, the iteration number needs to be about three times more than the area with a high signal level.