Vt1200 tissue slicer

Fresh pancreatic cancer tissue samples were procured immediately after surgical resection from University of Michigan Hospital. Informed consent was obtained from all patients. The remaining connective, fibrotic or adipose tissue were removed with razor blades. Tumor specimens were embedded in 3% low melting point agarose/PBS before cutting in the Leica VT1200 tissue slicer. The slice thickness ranged between 100-200 μm. Slices were then cultured in DMEM with penicillin (100 U/mL), streptomycin (100 U/mL) and amphotericin (Fungizone 2.5 μg/mL). All experiments were performed in triplicate and were repeated at least three times.

Hippocampal slices were prepared from 4–10 week old mice of either sex. Animals were anesthetized by isoflurane inhalation and euthanized. The mouse was decapitated, the brain removed and immersed for 5 min in ice-cold cutting solution (in mM: 125 NaCl, 4 KCl, 1.25 NaH₂PO₄, 26 NaHCO₃, 6 MgSO₄, 1 CaCl₂, and 10 glucose bubbled with 5% CO₂–95% O₂), and then mounted in a cutting chamber. Horizontal slices from the ventral half of the hippocampus were cut with a Leica VT1200 tissue slicer at 350–400 μm and allowed to recover for 45–60 minutes in artificial cerebrospinal fluid (aCSF; in mM, 125 NaCl, 4 KCl, 1.25 NaH₂PO₄, 26 NaHCO₃, 1.3 MgSO₄, 2.5 CaCl₂, and 10 glucose) containing 4 μM dipicrylamine (DPA, City Chemical LLC) at room temperature.