Arrangement and imaging of biological samples

摘要

High-resolution three-dimensional imaging of a specimen is facilitated. According to an example embodiment of the present invention, a series of very thin slices from a specimen are serially and robustly arranged on an imaging device such as a microscope slide. The slices are imaged and the images are used to reconstruct a three-dimensional image having high resolution at depths into the specimen. The serial arrangement of the slices facilitates the proper ordering of images for reconstruction. Further, the robust nature of the slice arrangement facilitates treatment of the slices and, in some applications, multiple treatments with corresponding imaging sequences for each treatment. Various embodiments are directed to methods and arrangements for three-dimensional characterization of biological specimen and to data that is accessible and/or executable by a computer for linking different images together in order to characterize such biological specimen in three dimensions.

主权项

1. A method for imaging a biological specimen, the method comprising:
embedding the specimen to set a molecular arrangement therein; thinly slicing the embedded specimen to provide a multitude of thin physical slices, having respective slice thicknesses defined between upper and lower surfaces of the respective slices of the specimen; serially arranging and fixing the physical slices in an array on a microscopy slide, therein forming an array of a multitude of slices; applying a stain to the physical slices; collecting image data from the slices on the slide, the image data including imaged portions of the specimen extending from the upper surface to the lower surface of each slice; using the image data and a known serial arrangement of the physical slices to combine image data from different slices to construct a three-dimensional image of the specimen; and after collecting image data, eluting the slices on the slide and repeating the steps of applying a stain and collecting image data with the eluted slices, wherein using the image data and a known serial arrangement of the slices to combine image data from different slices to construct a three-dimensional image of the specimen includes using image data from the slices before and after elution.