The shallow depth of focus in conventional light microscopy hinders the observation of the whole image when the object is thicker than the depth of field. Most of the existing techniques measured the object distance, which is not necessarily the actual distance of each pixel in the image. We implemented a means of determining the "best focus" of each pixel and located the height of object points by sectioning at different sample heights. Combining the height information and its gray values together, we obtained an image where the blur from the finite depth of focus is eliminated. Limitations of the technique are discussed together with composed images.ed images.
© 1997 Optical Society of KoreaPDF Article