Ex vivo applications of multiphoton microscopy in urology
2016
Background: Routine urological surgery frequently requires rapid on-site histopathological tissue evaluation either
during biopsy or intra-operative procedure. However, resected tissue needs to undergo processing, which is not only time
consuming but may also create artifacts hindering real-time tissue assessment. Likewise, pathologist often relies on
several ancillary methods, in addition to H&E to arrive at a definitive diagnosis. Although, helpful these techniques are
tedious and time consuming and often show overlapping results. Therefore, there is a need for an imaging tool that can
rapidly assess tissue in real-time at cellular level. Multiphoton microscopy (MPM) is one such technique that can
generate histology-quality images from fresh and fixed tissue solely based on their intrinsic autofluorescence emission,
without the need for tissue processing or staining.
Design: Fresh tissue sections (neoplastic and non-neoplastic) from biopsy and surgical specimens of bladder and kidney
were obtained. Unstained deparaffinized slides from biopsy of medical kidney disease and oncocytic renal neoplasms
were also obtained. MPM images were acquired using with an Olympus FluoView FV1000MPE system. After imaging,
fresh tissues were submitted for routine histopathology.
Results: Based on the architectural and cellular details of the tissue, MPM could characterize normal components of
bladder and kidney. Neoplastic tissue could be differentiated from non-neoplastic tissue and could be further classified as
per histopathological convention. Some of the tumors had unique MPM signatures not otherwise seen on H&E sections.
Various subtypes of glomerular lesions were identified as well as renal oncocytic neoplasms were differentiated on
unstained deparaffinized slides.
Conclusions: We envision MPM to become an integral part of regular diagnostic workflow for rapid assessment of
tissue. MPM can be used to evaluate the adequacy of biopsies and triage tissues for ancillary studies. It can also be used
as an adjunct to frozen section analysis for intra-operative margin assessment. Further, it can play an important role for
pathologist for guiding specimen grossing, selecting tissue for tumor banking and as a rapid ancillary diagnostic tool.
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