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Official websites use. Share sensitive information only on official, secure websites. This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4. Content: Multiphoton microscopy MPM is capable of producing optical sections of resolution that can match or surpass that of wide-field microscopy with significant advantages over slide-based approaches. Avoiding the need for wax-embedding, physical cutting, and separate staining, simplifies workflow and reduces costs and labor.
Digital images may become available in a shorter time frame. And the imaging is non-destructive and non-consuming, preserving tissue for ancillary studies. Optical sectioning also avoids challenges of slide imaging such as focus problems and cutting artifacts.
Perceived drawbacks of MPM include long sample processing times, long data-acquisition times, high cost, and image degradation with depth. We have sought to systematically address these limitations for the practical implementation of MPM in clinical diagnostics, a technique we call Clearing Histology with MultiPhoton microscopy CHiMP , and present preliminary data on its application to primary diagnosis of human prostate biopsies.
A custom multiphoton microscope based on a polygonal spinning mirror was designed and built for high speed and high resolution image capture at depth. Custom software tools were developed in python for image processing and visualization of multi-level image stacks. Design: Single prostate core biopsy specimens were obtained from consented individuals.
Processing goals were making images available same day of biopsy. Specimens were subsequently submitted for traditional physical slides and scanning on a WSI system.