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

expanders and collimators. The output from the solid state red laser is not a single line. Therefore, it is necessary to use an excitation filter to clean its output. A circular aperture is placed at the output of each Spatial Filter/Beam Expander, to select only the central portion of the laser intensity of a normally Gaussian intensity profile, so that the entrance pupil of the scan lens is filled with relatively uniform illumination and the maximum numerical aperture (resolution) of the lens is realized.

The three collimated lasers are made colinear with a beam combiner composed of a regular mirror and two different long pass dichroic mirrors. The regular mirror is used for deflecting the red laser to the combiner. One dichroic mirror reflects the shorter wavelength green laser and passes the red laser. Similarly, the other dichroic mirror reflects the blue and passes both the green and red lasers. The colinear laser beams are then reflected by the Primary Dichroic Beamsplitter and fed through the Beam Expander/Optical Focusing Mechanism, which is composed of two apochromatic lenses.

The Beam Expander/Optical Focusing Mechanism further enlarges the beam diameter so that it fills the entrance pupil of the scan lens, to maximize its numerical aperture, and hence the resolution limit. By controlled adjustment of the axial distance between the two beam expander lenses, the collimated laser beam can be tuned to be slightly diverging or converging, which helps to move the focal plane up and down along the optical axis of the scan lens, to bring thin specimens with different mounting media (e.g., coverslip) in focus. Optical sectioning can be realized by collecting stacks of images at different focal plane positions in thick specimens, which can be used for 3D reconstruction of structures.

2.3. Fluorescence Detection Optics

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