Optical sections by means of “structured illumination”: background and application in fluorescence microscopy
04.05.2006 - Photonik international /2006 | [ deutsche Version lesen ]
Fluorescence is a standard technique in modern biomedical research that is used for the specific detection of cell and tissue structures. A problem arises when “thick” specimens are used whose thickness in the direction of the optical axis is greater than the objective’s depth of field. In this instance, the sharp image information from the focal plane is overlaid with blurred image information from out-of-focus planes. As a result contrast and resolution in the z-direction are reduced and 3D reconstruction of the sample is inhibited. An “optical section” through the sample, which only extracts the information from the sample that is in focus, can prevent or even reverse this problem. One method for generating optical sections is to use “structured illumination”, as has been implemented in the ApoTome from Carl Zeiss.
Authors: Hubert Bauch, Jörg Schaffer, Carl Zeiss MicroImaging GmbH, Göttingen, Germany