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B. I. Gaynes, V. Mangal, C. Lo; Episcopic Imaging of the Mammalian Eye. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6365.
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© ARVO (1962-2015); The Authors (2016-present)
To illustrate the utility of a novel and near physiologic imaging system of transparent as well as non-transparent tissue through use of episcopic fluorescence image capture (EFIC) that preserves anatomical form-function relationships and presents them 3-dimensionally without distortion.
EFIC was conducted on intact murine globes and rabbit lenses by use of a Leica MZ16FA stereomicroscope mounted on the knife bridge of the microtome. This is an automated epifluorescence stereomicroscope with motorized focus and variable zoom, allowing multiple exposures of a single specimen at different magnifications using darkfield fluorescence imaging to visualize the tissue at the block surface via tissue fluorescence. Image capture was controlled with Openlab (Improvision, Inc.) automation using an Orca ER charged-coupled device (CCD). As the block is sectioned, the block face was sequentially imaged with the ORCA-ER camera, resulting in a serial image stack that is in perfect registration.
The resulting images provided high-resolution volumetric 3-D images of the mammalian eye that featured a haptic style viewing feature that allows visualization of tissue morphology from virtually any viewing angle. The attached figure illustrates a unidimensional replicate vector projection of the in-situ rabbit lens and ciliary body. The lens suture and lens fiber aggregates are discerned.
EFIC appears to offer significant novelty in terms of provision of an enhanced virtual imaging platform suitable for use as a virtual dissecting tool or in further embodiments, as a virtual microscopy instrument that features little if any distortion of tissue imagery.
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