Background: Measurement of muscle fiber size and determination of size distribution is important in the assessment of neuromuscular disease. Fiber size estimation by simple inspection is inaccurate and subjective. Manual segmentation and measurement are time-consuming and tedious. We therefore propose an automated image analysis method for objective, reproducible, and time-saving measurement of muscle fibers in routinely hematoxylineosin stained cryostat sections. Methods: The proposed segmentation technique makes use of recent advances in level set based segmentation, where classical edge based active contours are extended by region based cues, such as color and texture. Segmentation and measurement are performed fully automatically. Multiple morphometric parameters, i.e., cross sectional area, lesser diameter, and perimeter are assessed in a single pass.