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

ectives must acquire a large number of small images of the specimen, and then tile them together, which is tedious, inefficient and error-prone.

Microscopic imaging of large specimens is of considerable interest to life science, partly because of the high throughput efficiency it offers. However, there are only a few academic investigations of wide field-of-view microscopes reported elsewhere, e.g., microphotography with a thin white light slice,45 using single spatial frequency grid pattern,46 and objective lens array based large field-of-view imaging.47, 48 The sizes of specimen that can be imaged with conventional widefield microscopes are limited by the field-of-view of the microscope objectives. With the increase of power (magnification) and resolution (numerical aperture), the field-of-view becomes smaller and smaller. When using standard microscope objectives for scanning laser imaging, except for stage-stage scanning, in both the beam-beam and beam-stage scan methods, the scan size is limited by the field-of-view of the objective lenses. In many clinical studies, it is desirable to acquire images of whole diseased tissue sections, such as those of tumors, while retaining microscopic resolution.49

The wide field-of-view confocal microscopy imaging system reported here can be used for the examination of morphology and molecular biology images of clinical tissue specimens, and high throughput screening in drug discovery through efficient imaging of microarray slides. It is capable of operating in brightfield, reflection and epi- fluorescence imaging modes. A red, green and blue laser is used to produce fluorescence, brightfield and reflection images. This new confocal system makes fluorescence examination of large specimens with microscopic resolution possible for the first time without tiling. The wide field-of-view will help fully realize the high throughput po
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