Keywords: Confocal, drug discovery, fluorescence, imaging, microscope, microscopy, pathology, tissue, wide field-of-view, microarray
1.1. Optical Microscopy.
An optical microscope is an instrument designed to help observe fine details of the specimen. It must have three features, i.e., high magnification - producing a magnified image of the specimen; good resolution - distinguishing the neighboring fine details; and high contrast - making the details visible to the human eye or imaging device. The usefulness of any microscope depends not only on its ability to magnify, but more importantly, upon its ability to resolve details.
Optical microscopy refers to the techniques and applications of light microscopes, and can be classified as lens- based and lensless microscopy.1 The ultimate imaging performance of lens-based microscopy is limited physically by optical aberrations, wavelength and diffraction of light, and out-of-focus light in the image. Aberration corrections can be made to the microscope optics, more importantly the objectives; light diffraction sets the optical resolution limit; and out-of-focus light in the image degrades both the resolution and contrast. Two optical techniques are available to reduce the out-of-focus influence in the image: confocal microscopy2-8 which employs a point-like illumination and a detection pinhole aperture, and nonlinear microscopy including two photon and multiphoton fluorescence microscopy,9-15 and second and third harmonic generation.1 Near-field microscopy falls into the lensless category.1'"/>
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