CHROMATISM AND CONFOCALITY IN CONFOCAL MICROSCOPES

In confocal microscopes, whenever a broadband light source is used, or when excitation and detection are performed at different wavelengths, for example in fluorescence, then the influence of microscope objective chromatism on the degree of confocality is very important. With poorly corrected objectives, depth of field will be increased and in the case of fluorescence the image may be lost altogether. Presented here are observations with truly achromatic reflecting objectives and with the same objectives modified by introduction of a known amount of chromatic aberration. The results should encourage manufacturers to consider development and production of both reflecting microscope objectives and refractive lenses with more carefully designed/controlled chromatic aberration.