Real-Time Monitoring of Acupuncture-Induced Dynamics in Subcutaneous Connective Tissue via In Vivo Laser Confocal...

Real-time dynamic monitoring of acupuncture effects is crucial for elucidating its biological mechanisms, yet traditional histological methods fail to capture transient responses in the live microenvironment. Here, we established a methodological framework based on in vivo laser confocal imaging, enabling high-resolution, real-time observation of morphological changes in local tissues following acupuncture intervention. After intravenous injection of sodium fluorescein in rats, dynamic alterations in microcirculation and extracellular matrix (e.g., collagen fiber arrangement) were tracked during acupuncture. The results demonstrated that this method clearly distinguished immediate (e.g., vasodilation) and delayed (e.g., inflammatory response) tissue reactions induced by acupuncture, with a spatial resolution of 0.5 µm and a temporal resolution of 3.5 fps. Subsequent quantitative analysis enabled objective measurement of morphological parameters. This protocol, described in this study, provides a visualization tool for in vivo studies of acupuncture mechanisms and serves as a technical reference for evaluating the effects of other physical stimuli (e.g., moxibustion).