Multifaceted Brain Tissue Analysis

Mice were anesthetized and perfused intracardially with 0.9% saline. Knee and brain samples were collected and post-fixed for 12 h in 4% paraformaldehyde. Brain tissue sections of 20 μm thickness were acquired with a CM3050S microtome (Leica, Wetzlar, Germany). Brain tissue sections were permeabilized in 0.3% TritonX-100 containing phosphate-buffered saline (PBST), followed by blocking with 0.3% normal donkey serum and 1% bovine serum albumin for 60 min at room temperature. The sections were then incubated with ionized calcium binding adaptor molecule 1 (Iba1, microglia marker, 1:200, goat, Novus Biologicals, NB100-1028), β-amyloid (amyloid plaques marker, 1:200, mouse, Santa Cruz, sc-28365), or Neu-N (neuron marker, 1:1000, rabbit, Millipore, ABN78) as described previously [13 (link)]. Sections were mounted with DAPI (Molecular Probes, Life Technologies).