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Endometriosis is a common benign condition affecting 6%–10% of women, the underlying cause of which remains largely unknown. Laboratory animal models of endometriosis offer a translational approach to advance research into this condition. To this end, the provision of a noninvasive method to monitor real‐time disease progression and elucidate the pathophysiological mechanisms of endometriosis is essential. This review evaluates advanced preclinical imaging techniques for studying endometriotic lesions in murine models and comparing and highlighting their technical specifications, advantages, and limitations. Imaging methods are categorized into optical (bioluminescence, fluorescence, and nanoparticle‐assisted near‐infrared) and nonoptical (magnetic resonance imaging, ultrasound, and positron emission tomography) modalities. Bioluminescence imaging enables noninvasive monitoring of lesion growth and early angiogenesis in luciferase transgenic mice but is limited by low signal intensity in superficial lesions. Fluorescence imaging does not require substrate injection but cannot distinguish between viable and dead cells. Near‐infrared, enhanced by nanoparticle permeability, allows effective visualization of endometriotic lesions, although accumulation in nontarget tissues may compromise accuracy in superficial regions. Magnetic resonance imaging facilitates longitudinal studies of ectopic tissue growth, offering rapid data analysis with minimal stress to animals. High‐resolution ultrasound provides a noninvasive assessment of lesion dynamics but has low sensitivity for detecting small lesions. Positron emission tomography imaging shows promise for identifying small superficial lesions in humans through the use of radiopharmaceuticals. The effectiveness of these imaging techniques depends on lesion depth, with all methods performing better for deeper lesions than for superficial ones. Ongoing research aims to integrate multiple imaging modalities to improve the efficacy of preclinical studies.

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