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Abstract:
Multiphoton microscopies (MPM), including two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy, are robust methodologies for visualizing and assessing tissue in the context of injury and cancer. SHG, a nonlinear scattering phenomenon, offers sensitivity to structures lacking axial symmetry, with collagen—a crucial protein surrounding cells—being a prime example. Collagen plays a supporting role in wound healing and cancer progression, with alterations in its structure serving as indicative markers of both processes. This work delves into the versatility of multiphoton and SHG microscopy in analyzing collagen’s involvement in wound healing and disease development, while also exploring creative solutions to microscope design that enhance the viability of endoscopic implementation.
Specifically, MPM with TPEF and SHG is used to visualize and quantify the steps in Achilles tendon healing following mechanical and thermal injury in rats. Also, the utilization of randomly sampled SHG measurements in colorectal cancer diagnosis is proposed for the first time, potentially streamlining endoscope design complexities related to the scanning mechanism. Further, the incorporation of angle-resolved SHG measurements in assessing colon tissue holds promise in eliminating the necessity for paired normal and cancer-suspicious measurements in each patient.
While MPM is an established microscopy in research settings, this investigation into novel device designs and expanded applications points towards an intriguing new direction for this technology. With these advancements, MPM with SHG is moving closer to achieving simpler and more feasible clinical application.