Scanning electron microscopy (Hitachi-SU3500) was used to analyze the ultrastructure of the intestine. Fifteen broiler chickens were randomly sampled and necropsied. Intestine (jejunum) was taken and sliced 4 cm. The jejunum samples were prepared using the biological sample preparation method for scanning electron microscope (Titze & Christel, 2016) (link).
The preparation process of the intestinal sample includes a cleaning process involving soaking in a phosphate buffer pH 7 for 5 hours, followed by an agitation process in an ultrasonic cleaner for 7 min. The sample was prefixed into 2.5% glutaraldehyde solution for 24 hours and continued the fixation process in 2% tannic acids solution for 5 hours, then washed with phosphate buffer pH 7 for 15 min 4 times. The next process was the dehydration of the sample by soaking it in alcohol with graded concentrations (50, 70, 85, 95, and absolute). After passing through the dehydration process, the sample was dried by soaking in tetra-butanol, placed in the freezer until frozen, and then put into a freeze-drier for 24 hours. The prepared sample was subsequently placed on a stub specimen and coated with Au using an ion coater tool. Further, the length of jejunum villi was evaluated by using an image processing from a scanning electron microscope (Hitachi SU-3500).
The preparation process of the intestinal sample includes a cleaning process involving soaking in a phosphate buffer pH 7 for 5 hours, followed by an agitation process in an ultrasonic cleaner for 7 min. The sample was prefixed into 2.5% glutaraldehyde solution for 24 hours and continued the fixation process in 2% tannic acids solution for 5 hours, then washed with phosphate buffer pH 7 for 15 min 4 times. The next process was the dehydration of the sample by soaking it in alcohol with graded concentrations (50, 70, 85, 95, and absolute). After passing through the dehydration process, the sample was dried by soaking in tetra-butanol, placed in the freezer until frozen, and then put into a freeze-drier for 24 hours. The prepared sample was subsequently placed on a stub specimen and coated with Au using an ion coater tool. Further, the length of jejunum villi was evaluated by using an image processing from a scanning electron microscope (Hitachi SU-3500).