Gold seal rite on ultra frost

Fifteen-micron-thick cryostat sections were mounted onto non-polarized glass slides (Gold Seal Rite-On Ultra Frost, Thermo-Scientific), thawed, slightly fixed in 75% ethanol for 40 sec, and stained with Thioflavin-S 0.05% in 100% ethanol (ThioS, Sigma-Aldrich, Cat# 1326-12-1) for 5 min, followed by dehydration in increasing concentrations of ethanol and xylene, and air dried. Immediately, sections were placed onto the stage of an Arcturus Veritas LCM apparatus (Arcturus, Thermo-Scientific) and laser capture microdissection (LCM) was performed with a laser power of 80–85 μV and pulse duration of 3,500 μsec to fill up ten CapSure LCM Macro caps (Thermo-Scientific, Cat# LCM0211) per region of interest and per donor. In CTRL donors, approximately 100 mm² of cortex were dissected. In AD tissues, four different regions of interest were dissected: ThioS+ plaques (approx. 1,000/donor), the 50 μm area around those ThioS+ plaques, ThioS+ NFTs with the 50 μm area around them (approx. 1,500/donor), and cortex far (>50 μm) from both ThioS+ plaques and NFTs (approx. 80–100 mm²).

Fifteen-micron-thick cryostat sections were mounted onto non-polarized glass slides (Gold Seal Rite-On Ultra Frost, Thermo-Scientific), thawed, slightly fixed in 75% ethanol for 40 s, and stained with Thioflavin-S 0.05% in 100% ethanol (ThioS, Sigma-Aldrich, Cat# 1326-12-1) for 5 min, followed by dehydration in increasing concentrations of ethanol and xylene, and air dried. Immediately, sections were placed onto the stage of an Arcturus Veritas LCM apparatus (Arcturus, Thermo-Scientific) and laser capture microdissection (LCM) was performed with a laser power of 80–85 μV and pulse duration of 3500 μs to fill up 10 CapSure LCM Macro caps (Thermo-Scientific, Cat# LCM0211) per region of interest and per donor. In CTRL donors, approximately 100 mm² of cortex were dissected. In AD tissues, four different regions of interest were dissected: ThioS+ plaques (approximately 1000/donor), the 50 μm area around those ThioS+ plaques, ThioS+ NFTs with the 50 μm area around them (approximately 1500/donor), and cortex far (> 50 μm) from both ThioS+ plaques and NFTs (approximately 80–100 mm²).