Home | DTI | 2006–07 funded proposals | Peter A. Santi, James Leger, Curt Morrison
Peter A. Santi, James Leger, Curt Morrison
Development of a Thin-Sheet Laser Illuminator for Optical Sectioning
This project proposes to develop a thin-sheet, laser illumination device that can be used with a fluorescent light microscope in order to produce optical sections of thick tissues. A z-series stack of well-aligned images are necessary for 3D reconstruction of tissues to better understand their complex anatomical and functional relationships. This device will fill a niche between the relatively lower resolution microMRI and microCT, which is used for thick tissues and whole organs, and the higher resolution of confocal/ multiphoton microscopy, which can only be used for thin tissues. A device prototype was previously constructed using a cylindrical lens to produce the light sheet. However, its performance was not optimal and we propose to replace the cylindrical lens with a diffractive lens for improved resolution and contrast of the images. Construction and testing of the new device will be performed by faculty, staff and a graduate student in the Department of Computer Science/Engineering and the Nano Fabrication Center. Performance testing using a whole mouse cochlea will be performed by faculty and an undergraduate student in the Cochlear Anatomy Laboratory of the Department of Otolaryngology. Fitting of the device to a microscope and collection of images will be performed by a collaboration with staff from Leeds Precision Instruments, Inc. Leeds will provide staff time and an Olympus light microscope with a digital camera, and a motorized zstage in order to collect a stack images of the cochlea. This new, potentially patentable, thin-sheet laser illuminator should offer outstanding performance over published, but not yet commercially available technology for the efficient, cost effective, collection of high resolution, stack of images from thick tissues.
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