At 90 days of storage, bread crumb was submitted to environmental
scanning electron microscopy (ESEM), carried out by means
of the Inspect S50 electron microscope (FEI, Hillsboro, Oregon, USA)
operating at an accelerating voltage of 15 kV. Before testing, the
first layer of each sample was carefully removed. Then, the sample
(small portions of bread cut with razor blade) were prepared for
ESEM analysis. Bread samples were mounted on specific stubs using
a thin spatula and transferred to the microscopy support. Digital
images at 100 were acquired by using the Imix software
(Princeton Gamma Tech, Princeton, USA).