Understanding the innervation of nociceptive afferents in the stomach is pivotal for elucidating the mechanisms underlying pain perception and gastrointestinal disorders. Immunohistochemical labeling of common pain markers, including substance P (SP), calcitonin gene-related peptide (CGRP), and transient receptor potential vanilloid 1 (TRPV1), has been widely employed to visualize nociceptive afferents. However, existing studies predominantly rely on sectioned tissue, limiting the holistic assessment of axonal networks and presenting challenges in terms of time and labor. To address these limitations, we introduce an automated imaging and analysis pipeline tailored specifically for investigating nociceptive afferent innervation in flat-mount preparations of the stomach. This innovative pipeline combines advanced imaging techniques and sophisticated image analysis algorithms to facilitate precise quantification and characterization of nerve fibers. Through the implementation of this automated approach, we have successfully examined the distribution, density, and morphology of nociceptive afferent fibers in the stomach. Notably, our methodology enables the first-ever comprehensive imaging of the entire nociceptive afferent innervation in a rat stomach. Furthermore, we employed the advanced neuron tracing software, Neurolucida 360, to characterize spinal afferent fibers and to achieve full tracing of CGRP-IR axon fibers in the mouse stomach. This comprehensive tracing facilitates the identification of key structures involved in nociceptive processing within the gastric system. In the context of pain suppression, we propose an alternative approach by targeting the cooling of spinal dorsal root ganglia (DRG; the origin of nociceptive axons in the stomach). We've designed and fabricated a compact micro-liquid thermo-regulator (MLTR) device utilizing 3D printing technology to address this. Potentially, The MLTR device can be directly implanted into the rat DRG, enabling precise temperature control through microfluidic and thermo-regulation methods. In the future, we will test whether the MLTR device effectively modulates nociceptive transmission by lowering the local temperature, providing a potential alternative to opioid-based pain suppression.


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Graduation Date





Cheng, Zixi


Master of Science (M.S.)


College of Medicine


Burnett School of Biomedical Sciences

Degree Program



CFE0009751; DP0027859





Release Date

August 2028

Length of Campus-only Access

5 years

Access Status

Masters Thesis (Campus-only Access)


College of Medicine

Restricted to the UCF community until August 2028; it will then be open access.