Nis elements br version 4

As described above, mice were euthanized using CO₂ inhalation, followed by decapitation. Each brain collected was weighed, sagittally sectionized, and hemi-brain included in 1.5 mL of phosphate-buffered saline (PBS), then macerated initially using a 5ml syringe attached to a 18G hypodermic needle, making delicate movements (aspiration and disposal) until a homogenate is obtained. The process was repeated with a 21G hypodermic needle, to homogenize the smaller particles, until a homogenate was obtained. The number of cysts was determined by optical microscopy analyzing 20 μL of the homogenate in duplicate, and the total number calculated for the whole brain. The size of the cysts was determined using the digital software NIS Elements BR version 4.3 (Nikon Co., Japan), using images obtained with a Sight DS-U3 color vision digital camera adapted to an Eclipse Ci-S microscope. The diameter of the cysts was measured with the digital morphometric apparatus NIS Elements BR version 4.3 software (Nikon Co., Japan). Through a Sight DS-U3 color-view digital camera adapted to an Eclipse Ci-S microscope. The frequency distribution of the diameter length of the cysts was grouped into classes and the number of occurrences in each class was counted to know the behavior related to the size of the cysts, throughout the infection.

Hemoxylin-eosin-stained sections of 10 well-oriented villi and crypts of duodenum, ileum and caecum from each animal were photographed with a 10× lens using a light microscope (Eclipse Ni-U; Nikon, Tokyo, Japan) and used to measure the villus height (VH) and the crypt depth (CD). The total number of goblet cells per villus, the number of goblet cells with different types of mucins as distinguished by conventional and lectin histochemistry were determined by counting of 10 well-oriented duodenum, ileum and caecum sections. The fluorescence signal intensity of stained GCs and no-stained epithelial cells (selected as background) were measured with the image-analyzing program NIS Elements BR (Version 4.30) (Nikon, Tokyo, Japan). The fluorescence intensity of GCs was calculated after subtraction of the background signal, and the mean intensity value was computed.
Values were expressed as means ± standard error (SE). The results were evaluated for statistical significance by Student’s t test.