When recording three-dimensional (3D) images by the method of optical sectioning microscopy, each optical section contains the in-focus information plus out-of-focus contributions that obscure the in-focus detail and reduce contrast. There are several methods to remove or prevent the out-of-focus contributions from the stack of optical sections. One such method is image estimation -the use of a computer program based on a mathematical description of the microscope to remove the out-of-focus contributions. Another method is the use of structured illumination and a simple arithmetic operation to obtain a image that in which the out-of-focus contributions are greatly reduced. We derived a method for image estimation that uses the images collected from the structured-illumination microscope. The method improves the resolution of small detail over that possible with the structured illumination using the simple arithmetic formula.

Date Published: 24 March 2005
PDF: 8 pages
Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005); doi: 10.1117/12.602633

Published in SPIE Proceedings Vol. 5701:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)