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

ign="left">Fluorescence microscopy provides many opportunities to understand biological structures and processes, such as the possibility of selective excitation in a mixture of different molecular species, the high sensitivity which allows a relatively small number of fluorescence molecules to be detected, with the availability of a variety of fluorescence indicators, and the ability of imaging fluorescence reporter proteins.54 In addition, the advantages of higher sensitivity, optical sectioning, and 3D analysis of images available due to confocal microscopy provide more information about biological objects.54

For fluorescence imaging, the fluorescence emission from the specimen is collected by the laser scan lens, while the raster scanning (beam scanning and stage scanning) is taking place, and the detected fluorescence signal is de-scanned simultaneously by the scanning mirror. With its diameter reduced by the backward beam expander, the fluorescence light passes the Primary Dichroic Beamsplitter, gets reflected by one of the detection dichroic filters or the detection mirror depending on the wavelengths of the fluorescence band, and passes through the respective emission filter on the filter wheel, before it is focused with the detection lens (achromatic focusing lens, not shown) and arrives at the detection pinhole (not shown) in front of the detector, which helps to reject out-of-focus light so that the confocal effect is realized. Due to the confocal effect, only the light signal from the detection volume located in the focal plane of the specimen can reach the photon detector. The dichroic filters in the detection path are long pass filters, which reflect light of shorter wavelengths and pass longer wavelengths.

The respective emission filters (barrier filters) on the filter wheel stop excitation light from entering the detectors, so that only the fluorescence signals are detected
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