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Abstract

Systems and methods are provided for staining tissue with multiple biologically specific heavy metal stains and then performing X-ray imaging, either in projection or tomography modes, using either a plurality of illumination energies or an energy sensitive detection scheme. The resulting energy-weighted measurements can then be used to decompose the resulting images into quantitative images of the distribution of stains. The decomposed images may be false-colored and recombined to make virtual X-ray histology images. The techniques thereby allow for effective differentiation between two or more X-ray dyes, which had previously been unattainable in 3D imaging, particularly 3D imaging of features at the micron resolution scale. While techniques are described in certain example implementations, such as with microtomography, the techniques are scalable to larger fields of view, allowing for use in 3D color, X-ray virtual histology of pathology specimens.

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