Ex vivo Autoradiography of 64Cu-TIMP2 in Mouse Brain

At 24 h after injection of ⁶⁴Cu-labelled TIMP2 (7.57 ± 0.15 MBq), mice were deeply anaesthetized with 2.5% (v/v) Avertin and perfused with ~30 ml chilled PBS. Brain tissue was quickly embedded in optimal-cutting temperature compound (Tissue-Tek), and coronal or sagittal sections (20 μm) were obtained for ex vivo autoradiography. Autoradiography was conducted using previously described methods^{49 (link)}, confirming anatomical localization by Nissl staining (cresyl violet acetate; Sigma Aldrich) using standard techniques. In brief, sections of 20 μm were mounted on microscope slides (Fisherbrand Superfrost Plus Microscope Slides), air-dried for 10 min, and then exposed to a digital storage phosphor screen (Perkin Elmer, USA) for 72 h at −20 °C. The image plate was analysed using a Typhoon 9410 Variable Mode Imager (Amersham Biosciences, USA) and images were visualized and processed by ImageJ (image processing and analysis software in Java, version 1.45s).