Purpose: Machine vision-based image processing methods can be useful for estimating the fresh weight of plants. This study analyzes the ability of two different image processing methods, i.e., morphological and pixel-value analysis methods, to measure the fresh weight of lettuce grown in a closed hydroponic system. Methods: Polynomial calibration models are developed to relate the number of pixels in images of leaf areas determined by the image processing methods to actual fresh weights of lettuce measured with a digital scale. The study analyzes the ability of the machine vision-based calibration models to predict the fresh weights of lettuce. Results: The coefficients of determination (> 0.93) and standard error of prediction (SEP) values (< 5 g) generated by the two developed models imply that the image processing methods could accurately estimate the fresh weight of each lettuce plant during its growing stage. Conclusions: The results demonstrate that the growing status of a lettuce plant can be estimated using leaf images and regression equations. This shows that a machine vision system installed on a plant growing bed can potentially be used to determine optimal harvest timings for efficient plant growth management.