This study investigated strain differences within the patellar tendon (PT) mid-region using an ultrasound-based digital image correlation (DIC) method. Six healthy young participants performed six knee extensions to 60% of maximal voluntary isometric contraction on 2 days. Sagittal ultrasound videos recorded during each contraction were analyzed using the DIC method to determine the strain-torque relationships of the superficial, deep, and whole PT mid-regions. Significantly greater strain was observed in the deep vs superficial layer of the PT mid-region for all contraction intensities, with peak strains of 5.8% (SD 1.7) and 4.5% (SD 1.5), respectively. DIC-based measures of peak tendon strain were repeatable within [intraclass coefficients (ICC) >0.97] and between sessions (ICCs >0.83) and agreed well with the conventional point-to-point method. This study confirmed that significant differences exist between deep and superficial layers of PT mid-region during ramped isometric extensions. These findings support the use of DIC to examine regional strain patterns within the PT mid-region that may be important in the context of tendon injury and adaptation.