title: SC-ZF-Hypo5
design: we worked swiftly with cold, well-oxygenated solutions, optimizing Papain incubation times for tissue age and brain region. We used caution with Ca2+ and Mg2+ in aCSF to avoid RNA extraction/RT interference, favoring Ca2+/Mg2+-free aCSF if not diluted downstream. Modified aCSF had specific component concentrations, and our materials included essential equipment and anesthesia. We prepared aCSF from an 8x stock with adjusted CaCl2 and MgSO4 concentrations, kept it fresh, and oxygenated it. The zebrafish procedure began with lethal anesthesia, brain dissection in ice-cold aCSF, embedding in 1.5% gel, and vibratome sectioning. Tissue was dissociated in the Papain vial, triturated, filtered, and washed, followed by centrifugation. The pellet was resuspended in aCSF with DNase, assessed for cell viability, counted, and diluted for use.