Project 1: The Mouse Brain Library Project 2: Internet Microscopy (iScope) Project 3: Neurocartographer and Segmentation of the MBL Project 4: The Neurogenetics Tool Box


















Principal Investigator/Program Director Williams,Robert W.

  Compromises and cost.

The design of the microscope and video equipment is inevitably a compromise between cost, ease of implementation, and image quality. For example, use of oil-immersion objectives with high numerical aperture would improve resolution from 0.8 to 0.25 µm, but for obvious reasons, oil immersion is not practical for an automated slide-handling system. We believe that for streaming video and even for acquiring stacks of images for 3D reconstruction, our best option is a dry objective with a moderately high numerical aperture (NA of 0.60.8) and a moderately long working distance. We now use an objective that can tolerate some variation in cover glass thickness without suffering serious spherical aberration. An alternative would be to use microscopes with motorized nosepieces and multiple objectives, but besides being far more expensive, these options would complicate the implementation without greatly extending the utility of the MBL. To provide a broader range of magnification, we will use a beam splitter and two video cameras each equipped with a different intermediate lens on microscopes designed for streaming real-time video. We can readily achieve a 3:1 ratio of magnification from two cameras using a single 40x objective. With this equipment, microscopists will have access to both high- and low-magnification video streams simultaneously, and they will of course have access to the 25 and 4.5 µm images for all cases in the library.

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  Streaming Video.