A single-cell transcriptomic atlas reveals a new myeloid parenchymal population in the zebrafish brain
A single-cell transcriptomic atlas reveals a new myeloid parenchymal population in the zebrafish brain
FACS-sorting of cd45:Dsred cells. Brains from adult Tg(p2ry12:GFP; cd45:DsRed) fish (n=3), were dissected in 0.9X Dulbecco′s Phosphate Buffered Saline (DPBS) were triturated and treated with Liberase TM at 33°C for 30-45 minutes, fully dissociated using a syringe with a 26G needle and washed in 2% fetal bovine serum diluted in 0.9X DPBS. Cell suspensions were centrifuged at 290g 4ºC 10 min and filtered through a 40µm nylon mesh. Just before flow cytometry analysis, using calcein violet to exclude dead cells (1µM, Thermo Fisher). Flow cytometry acquisition and cell sorting was performed on a FACS ARIA II (Becton Dickinson).  RNA Illumina