<jats:title>Abstract</jats:title>
          <jats:p>Advanced imaging tools are revolutionizing the diagnosis, treatment, and monitoring of medical conditions, offering unprecedented insights into live cell behavior and biophysical markers. We introduce a modular, hand-held fluorescent microscope featuring rapid set-up and sub-millimeter resolution for real-time biological analysis. We apply our system to map pH and nitric oxide (NO), biomarkers central to wound healing, in subcutaneous wounds. Using machine learning to cluster pH reveals spatiotemporal trends, including a concentric gradient peaking at the center and stabilization at the wound edge. NO clustering shows high-concentration structures that decrease in size but intensify as healing progresses from hemostasis to proliferation, enabling prediction of the healing day and re-epithelialization. These biomarker mappings offer insights poised to inform future wound healing studies. This research lays the groundwork for integrating the modular imaging unit with bioelectronic devices in closed-loop feedback systems, using machine learning to guide optimal wound treatment and accelerate healing.</jats:p>