3D Light Microscopy Images To Aid in Representation

Over the years 3D images have been both desired and used to provide an enhanced view of microscopic specimens, so as to provide or convey a better perspective for the viewer. In general, one can say there are two approaches to 3D representation, both tied to human factors. The first approach is based on stereographic image production and the second is based on serial or continuous depth of focus. The stereographic approach relies on the natural human perspective of viewing an object from two eyes gathering light waves at offset angles from one another that are then combined into a single image by the brain. The 3D effect derives from angle of view to a fixed focal plane and the space within the depth of focus range for that plane. The second approach attempts to create an image completely in focus in all planes. The first can be achieved by tilting the specimen, using offset optics to mimic stereography, using color separation and isolation techniques, and modifying the optical path in two different ways. The second approach typically uses image stacking but could be accomplished with continuous selective directed lighting.