Multi-Spectral Three Dimensional Imaging System and Method

A computer-implemented method of creating an image of a specimen wherein the specimen is contacted with at least one first fluorophore having an emission spectrum in the range of approximately 200 nm to 1000 nm, the method comprising the steps of:

• i. receiving, by a processor, a first image Nx of a first section Sx of a specimen, the first image created using a first wavelength of invisible light;
• ii. receiving, by the processor, a second image Nx+1 of a second section Sx+1 of the specimen, the second section Sx+1 being adjacent to the first section Sx and the second image created using the first wavelength of invisible light;
• iii. co-registering, by the processor, the first image and the second image;
• iv. creating, by the processor, a single-plane image of the first section using a next-image process;
• wherein the creating of the image includes a debluring process;
• v. receiving, by the processor, an image Nx+3 of a third section Sx+3 of the specimen created using the first wavelength of invisible light;
• and,
• vi. creating, by the processor, a 3D image of the specimen using at least three images of adjacent specimen sections.