Water-holding capacity (WHC) is a primary quality determinant of salmon flesh. One of the limiting factors for not having a direct measurement of WHC for salmon quality grading is that current WHC measurements are destructive, time-consuming, and inefficient. In this study, two hyperspectral image systems operated in the visible and short-wave near infrared range (400-1000 nm) and the long-wave near infrared range (897-1753 nm) were applied for non-invasive determination of four WHC indices, namely percentage liquid loss (PLL), percentage water loss (PWL), percentage fat loss (PFL), and percentage water remained (PWR) of salmon flesh. Two calibration methods of partial least square regression (PLSR) and least-squares support vector machines (LS-SVM) were applied, respectively, to establish calibration models of WHC indices based on the spectral signatures of salmon flesh, and the performances of these two methods were compared to determine the optimal spectral calibration strategy. The performances were also compared between two hyperspectral image systems, when full range spectra were considered. Out of 121 wavelength variables, only thirteen (PLL), twelve (PWL), nine (PFL), and twelve variables (PWR) were selected as important variables by using competitive adaptive reweighted sampling (CARS) algorithm to reduce redundancy and collinearity of hyperspectral images. The CARS-PLSR combination was identified as the optimal method to calibrate the prediction models for WHC determination, resulting in good correlation coefficient of prediction (rP) of 0.941, 0.937, 0.815, and 0.970 for PLL, PWL, PFL, and PWR analysis, respectively. CARS-PLSR equations were obtained according to the regression coefficients of the CARS-PLSR models and were transferred to each pixel in the image for visualizing WHC indices in all portions of the salmon fillet. The overall results show that the laborious, time-consuming, and destructive traditional techniques could be replaced by hyperspectral imaging to provide a rapid and non-invasive measurement of WHC distribution in salmon flesh.