Application of epifluorescence light microscopy (EFLM) to study the microstructure of wheat dough: a comparison with confocal scanning laser microscopy (CSLM) technique

To study dough microstructure, epifluorescence light microscopy (EFLM) combined with digital image processing software was used, which enabled an improved image quality. A comparison was made between EFLM and confocal scanning laser microscopy (CSLM) methods. Both techniques were satisfactorily able to demonstrate changes in the dough microstructure upon different stages of z-blade mixing. Dough mixed for a shorter time (under-mixed) showed a heterogeneous structure with coarse protein domains and clusters of starch due to local segregation or de-mixing effect. Increasing mixing time (optimal mixing) led to development of interconnected gluten network covering starch granules throughout the dough, representing optimal development. Over-mixing led to formation of a homogeneous dough microstructure in which the gluten phase showed a fine distribution throughout the dough. Using a double staining method in the preparation of samples for both microscopic techniques it was possible to observe gluten network structures together with starch granules. Moreover, special features of image processing software described in this study enabled us to improve EFLM images and to obtain comparable images with CSLM. This could favour a low cost and a convenient microscopic observation of biomaterials.