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Experimental strategies to stimulate axon regeneration are the core of therapeutic development for nervous system injury and disease. Here, we present AxoMetric, an open-source, user-friendly software for rapid and automated quantification of regenerated axons in nerve tissue sections. AxoMetric automatically identifies regenerated axons, providing quantitative measurements at customizable distances from the injury site that closely match conventional manual quantification but with greater speed and consistency. Additional features of AxoMetric include quantification of neuronal cell bodies, normalization of axon density to nerve diameter to account for anatomical variability, and an image-queuing function for high-throughput analysis. We demonstrate its application to quantify regenerated axons in the optic nerve, cell density in retinal flatmounts, and regenerated axon bundles in injured sciatic nerves. Collectively, AxoMetric streamlines and standardizes quantitative assessments of neural regeneration across diverse experimental paradigms and laboratories.