Fluorescence and Confocal Microscope: Basic Principles and Applications in Pathology

Abstract

Fluorescence microscope applies high-intensity light to illuminate the substance that emits fluorescence light. The present chapter describes the basic principle and applications of fluorescence microscopy. There are two types of fluorescence microscope: transmitted fluorescent microscope and incident fluorescent microscope. The working principles of both these types of microscopes are described. The confocal fluorescence microscopy (CFM) provides three-dimensional optical resolution. In CFM, at one time we see the image of the particular depth of the object at a small point. All the out-of-focus light is eliminated by passing the light through the pinhole. Multiple images at different depth are accumulated and then reconstructed to provide a three-dimensional image. The working principle and applications of CFM are discussed in the present chapter.