The aim of this study was to evaluate the capacity of two strains of lactic acid bacteria (LAB), Lactobacillus rhamnosus and Lactococcus lactis, and a yeast strain, Saccharomyces cerevisiae, inactivated by heat (121 °C, 10 min), from binding to aflatoxin M 1 (AFM 1 ), as well as the interaction between these microorganisms, aflatoxin M 1 and the Minas Frescal cheese matrix after 2 and 30 days of storage. The ability of LABs and S. cerevisiae to bind AFM 1 to Minas Frescal cheese was evaluated by high performance liquid chromatography (HPLC) composed of a fluorescence detector. The interaction between these microorganisms and AFM 1 was evaluated using a scanning electron microscope composed of a backscattered electron detector with a voltage of 15 kV and magnifications of 1000 ×, 5000 × and 8000 ×. The use of microorganisms as a biological method is efficient in reducing AFM 1 in Minas Frescal cheese and does not affect the microbiological parameters. AFM 1 reduction varied according to the microorganism used in the treatments. S cerevisiae showed greater capacity to bind AFM 1 over time, compared to LABs. Scanning electron microscopy was especially useful, confirming that lactic acid bacteria and S. cerevisae were able to bind AFM 1 particles in Minas Frescal cheese.