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

de field-of-view confocal system presented in here, the reflectance imaging mode shares the same illumination and detection optics with the fluorescence mode. Neutral density filter attenuation of laser power at the detection path is used for reflectance imaging, in place of the fluorescence emission filters. In reflectance imaging, the laser light reflected/scattered from the detection volume of the specimen is collected by the laser scan lens, descanned by the scanning mirror, and detected by the same confocal detector used in fluorescence imaging (with appropriate filter changes).

2.5. Transmission Imaging

Transmission (brightfield) microscopy is the most commonly used method in the examination of histological samples, based on the absorption of the illumination light by structures in the specimen. In a conventional widefield microscope, the commonly used light sources are tungsten filament lamps with a continuum of blackbody radiation, or xenon arc lamps with a continuum between 300 and 800 nm.21 However, in the brightfield detection mode of the wide field-of-view imaging system presented in this paper, the red, green and blue (RGB) lasers are used as the illumination source to focus simultaneously through the specimen. RGB imaging is based on the color perception theory. In general, a mixture of relative amounts of red, green and blue light can present most colors to the human eye. For example, the perception of yellow (e.g., 560 nm) can be evoked by either the presence of that wavelength or the presence of comparable intensities of red (600 nm) and green (530 nm). Broadly speaking, when transilluminated as in brightfield microscopy, if a sample appears green, the red and blue lights are absorbed.

As discussed in Sect. 1.1.1, in brightfield microscopy, the tissue sections are usually stained with one or more chromophores
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