Neurite degeneration is the process in which the axons and dendrites of a neuron undergo deterioration and is associated with the progression of various neurodegenerative diseases such as Parkinson’s disease (PD). Further studies are needed to understand the molecular factors that underlie neurite degeneration. One commonly used method of quantifying neurite degeneration involves analyzing micrographs using the software FIJI to calculate a Degeneration Index (DI) by determining the ratio of fragmented neurite area to total neurite area . However, this method can be time consuming and subject to inaccuracies. Here we set out to create a more efficient and accurate method of quantifying neurite degeneration. To model neurite degeneration associated with PD, we cultured Lund Human Mesencephalic (LUHMES) cells and induced oxidative stress in culture using 6-hydroxydopamine . Micrographs of degenerating neurites were then used to develop an enhanced method for measuring neurite degeneration. Through optimization of parameters for detecting neurite fragments, enhancing contrast of the neurite images, and using image processing to remove non-neurite materials from the image, we developed a method that calculates a more accurate DI. Additionally, we have created a macro to automate DI measurements, making the method more time efficient and user-friendly than previous methods. To exemplify an application of the improved method, we applied the method in experiments revealing the effects of c-Jun N-terminal Kinase signaling on oxidative stress-induced neurite degeneration. This automated method will allow investigators to more effectively assess neurite degeneration, an important, early-stage event associated with a variety of neurodegenerative diseases.