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A new wide field-of-view confocal imaging system and its applications in drug discovery and pathology

p> Computer image processing techniques can also be used to enhance the image quality, for example, deconvolution analysis of conventional widefield images to remove out-of-focus blur.

Two types of optical microscopes are in common use today, i.e., conventional widefield microscopes16-18 and the more recent confocal microscopes.2-8 Both of these have relative different advantages and disadvan- tages.6, 19, 20

1.1.1. Conventional widefield light microscopy

The name of widefield microscopy comes from the fact that the entire field of view is uniformly illuminated and observed. In both brightfield (transmission) or epi-fluorescence imaging, the specimen needs to be suficiently thin (e.g., 2 - 5 μm), so that structures at difeerent sections can be clearly observed to obtain maximum resolution. Absorption difference by the structures as light passes through the specimen and fluorescence emission difeerence from the structures when excited produce contrasts that reveals the details.

In widefield light microscopy, specimen staining is a routine process. Most living biological specimens are virtually transparent, and often remain so after fixation, especially after being sliced into thin sections.21 Imaging contrast can be realized by treating them with natural or synthetic dyes, a process called staining.21, 22 The commonly used hematoxylin and eosin staining colors the nuclei of cells blue-purple and other tissue components pink.23, 24 Generally, dyes are stains used in brightfield microscopy, and fluorophores in fluorescence microscopy. The term "stain" refers to both dyes and fluorophores.23 A dye is an organic aromatic molecule containing the requisite groups that provide visible color and permit molecular binding to a material. A
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