The effects of an osmotic dehydration process using sucrose and maltose solutions at 40 and 60 °Brix on microscopic features and some mechanical and thermal properties of guava tissue were studied. Also the addition of calcium lactate to the sugar solutions, aiming at preserving the structure of the processed fruits, was investigated. The guava texture (stress at failure) and the structure as observed by light microscopy were both evaluated, and differential scanning calorimetry (DSC) was used to verify the interaction between calcium ions and cell wall pectin in the guava tissue. The calcium content of the differently treated samples was also related to microscopic features, mechanical and thermal properties of guavas. The osmotic process using sucrose and maltose solutions caused severe structural damage to the guava tissue, and this effect was intensified at higher sugar concentrations and by the use of sucrose solutions. The addition of calcium lactate promoted maintenance of the guava structure, showing turgid cells with well-defined cellular contours, resulting in an increase in hardness and indicating bonding between the Ca2+ and cell wall pectin, which was confirmed by the DSC experiments.