title: single cell sequencing of chicken hindbrain
design: HH4 embryos electroporated with FoxD3-NC2:eGFP were cultured until HH12 ex ovo at 37C, following which the hindbrain region spanning rhombomeres 6, 7, and 8 was dissected under a fluorescence microscope. We pooled the dissected tissue washed in chilled 1X DPBS and incubated it in Accumax cell dissociation solution (EMD Millipore) for 15 minutes at 37C with gentle pipetting every 5 minutes. Dissociation was terminated using Hanks Buffered Saline Solution (HBSS) (Corning) supplemented with BSA Fraction V (Sigma; 0.2% w/v). The suspension was centrifuged at 300g for 4 minutes to collect cells at the bottom, the supernatant was removed, and the pellet was resuspended in 1mL HBSS-BSA. To remove cell debris and clumps, the 1mL suspension was passed through a 20m filter in a clean hood. This suspension was loaded on a 10X Chromium chip A (v2) to generate GEMs.