A new method for surface profiling in the confocal microscope utilizing chromatic aberration is presented. An expression for the integrated spectral intensity in the new profilometer is derived and chromatic aberration in the confocal microscope is explained. Estimates of the minimum resolvable depth and the maximum depth that can be profiled are given utilizing a ratiometric signal detection scheme. Examples of ratio images illustrating the new surface profiling technique are presented, showing the scheme to be both simple to implement and effective.
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.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.