title: single cell RNA-Seq of zebrafish: whole organism of juvenile
design: A single Juvenile fish at about 30 dpf was anesthetized in tricaine and euthanized on ice. Then it was washed three times with 200 L ice-cold HBSS supplemented with 1% BSA, and transferred into 200 L Dissociation buffer, 1  TrypLE (Thermo Fisher Scientific) in 1  HBSS [Ca+/Mg+ free] (GIBCO). Next, the solution was incubated for 20 minutes and mixed occasionally with a pipette. 120 L HBSS and 80 L Digestion cocktail stock (5 mg/mL collagenase (Sigma-Aldrich), 10 mg/mL pronase (Sigma-Aldrich), 10 mg/mL proteinase K (Sigma-Aldrich)) were added, and the suspension was pipetted until the larva was invisible. The cell suspension was washed with wash buffer twice and pelleted by centrifuging at 600g for 8 minutes at 4 C and resuspended in wash buffer. The cells were filtered through a 35-m strainer and processed using the 10x Genomics platform with Chromium Single Cell 3 v2 Reagent Kit.