Fat is of the most important ingredients in bakery products which contributes in providing textural properties of the baking products, particularly biscuits. Three entirely different olein (canola oil), palmitic (PBF) and lauric (PKBF) based fats were applied to the hard dough biscuits (HDBs) and their structural characteristics were evaluated. Results showed that significant differences (p < 0.05) were found in the outcome of the tests. Confocal laser scanning colorimetry (CLSM) of the biscuit doughs showed the highest and the least homogenous distribution of fats particles PBF and canola oil-based treatments. Moreover, experiments on biscuits doughs revealed that the PBF treated doughs had the most lowering effect on the hardness (156.37 ± 14.52) and springiness (0.91 ± 0.02), while it increased the cohesiveness (0.57 ± 0.02) and adhesiveness (−641.37 ± 29.41). Physical tests on biscuits were also in the same direction as the doughs except for browning index (BI) which was the highest in PKBF (26.98). Moreover, image processing on scanning electron microscopy (SEM) images showed the highest porosity and expansion formation was related to the PBF treatment, while canola oil treated biscuit highest fractal parameters which define the highest porosity irregularities in the biscuit structure.