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

to add contrast to such sections. The end result is to produce an amplitude object from a transparent phase object, so that the illumination light is absorbed differently across the specimen. The three lasers have wavelength lines of 633 nm (red), 535 nm (green) and 488 nm (blue). The common dyes used for tissue staining are mostly broadband dyes. Thus their absorption spectra will overlap with one or more laser lines, which makes RGB imaging using the three laser sources possible.

In confocal reflectance and epi-fluorescence imaging, the signal light is descanned by the scanning mirror so that the detection aperture tracks the focused laser spot at the specimen, and the full advantage of a confocal system, i.e., the two way rejection of out-of-focus light, is realized. In a confocal transmission configuration, however, the descanning is more technically challenging. The published solutions include the use of a second synchronized scanning system,2, 3 a complicated beam path to redirect the transmitted light back to the scan- ner,56 and the use of a nonlinear crystal as confocal pinhole for the detection of transmitted signals through second-harmonic generation without descanning.57, 58

The transmission imaging mode in the wide field-of-view system described in this paper does not implement a confocal pinhole, the transmitted light is simply detected using an RGB detector. However, the point illumina- tion of the specimen in the scanning system offers theoretically better signal-to-noise ratio than a conventional widefield microscope. The point-illumination scanning scheme does reduce the lateral background, and reduces photobleaching in the specimen. As the brightfield mode is not implemented in a confocal configuration, it is only suitable for applications in thin and stained tissue imaging.

3. APPLICATIONS IN DRUG DISCO
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