Laminin-Coated Coverslips for Cell Culture

Cells were plated onto laminin-coated (Sigma, L2020, 3.3 μg/mL) coverslips which had been treated with the self-assembled monolayer, N-1(3-[trimethoxysilyl]propyl)-diethylenetriamine (DETA, United Chemical Technologies Inc., Bristol, PA, T2910KG). Briefly, plasma-cleaned glass coverslips (Thomas Scientific 1217N81, 18 mm, no. 1.5) were silanized with a 0.1% (v/v) mixture of DETA in freshly distilled toluene (Fisher T2904) as described previously⁷³ . The DETA-treated glass was heated to just below the boiling point of toluene (70°C), rinsed with toluene, reheated to 70°C, and then dried in an oven overnight at (100–105°C). Surfaces were validated via contact angle measurement and X-ray photoelectron spectroscopy as described previously^{74 (link)}. Laminin was added to DETA-treated glass overnight at 4°C. DETA has previously been used for long-term culture of multiple cell types, including neurons^{72 (link),74 (link)–77} and enhances the adsorption of laminin onto the culture surfaces (see Supplemental Figure 1).