Figure 1

Preparation of hamster tissue slices. To obtain intestinal slices (upper panel), all the steps were performed under aseptic conditions, in no more than 1 h. (a) After dissection, the intestinal content was washed with cold KB buffer and the colon was divided in 3 cm segments; clean fragments were placed into ice-cold KB buffer. (b) The adhering fatty tissue was carefully removed. (c) One side of the intestinal fragments were tied with surgical thread and filled with 3% low melting point agarose at 37 °C. (d) Agarose infiltrated fragments filled with agarose were transferred into ice-cold KB buffer to permit the agarose solidification. (e) The intestinal segments were collocated in the tissue embedding unit of the Krumdieck tissue slicer and a pin was placed in the solid agarose to help the filling with 2% low melting point agarose at 37 °C of the mold in order to form cylinders containing the colon segments. (f) Colon tissue slices (300–400 µm thickness) were prepared using the Krumdieck tissue slicer and were collected in ice-cold KB buffer and incubated in culture medium at 37 °C. The process to obtain liver slices (lower panel) is simpler than intestinal slices; the process was also performed under sterile conditions, and in the shortest possible time. (g) After the liver was removed, it was placed into ice-cold KB buffer and the hepatic lobes were separated with a scalpel. (h) The hepatic lobes were cored into 5 mm diameter cylinders. (i) The cores were then placed in oxygenated KB buffer (4 °C, 95:5 O₂: CO₂). (j) Liver tissue slices with 250–300 µm thickness were prepared using the Krumdieck tissue slicer and were gently placed and incubated onto 24-well microplates with culture medium at 37 °C.