Subject Area:	•Biochemistry, Genetics and Molecular Biology •Medicine and Dentistry
More specific subject area:	Anatomy and Molecular Embryology
Method name:	Cell injection in CPM Bead implantation in the BA2 Whole-mount immunostaining for QCPN
Name and reference of original method:	Kodo K, Shibata S, Miyagawa-Tomita S, Ong SG, Takahashi H, Kume T, Okano H, Matsuoka R, Yamagishi H. 2017. Regulation of Sema3c and the Interaction between Cardiac Neural Crest and Second Heart Field during Outflow Tract Development. Scientific reports 7.
Resource availability:	A. Resources needed to reproduce the experiments Chicken eggs Quail cells (QT6) Fridge Incubator Tungsten needle Forceps Spoon Medical tape Fine scissors Falcon tubes Pasteur pipettes Eppendorf pipettes Caps of microcentrifuge tube Petri dish 24 Well plates Cellulose compress (900-0853, Henry Schein) Parafilm 38 M X 10 CM Aspiratory tube Glass capillary Stereo microscope (M165 FC, Leica, Germany) equipped with a digital camera (DFC420 C, Leica, Germany) PBS (1x):Start with 800 ml of distilled water, 8 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, 0.24 g of KH2PO4, adjust the Ph to 7.4 with HCl, add distilled water to total volume of 1 L. PBST:0.1% Tween-20 in 1x PBS. 4 % PFA in PBS:for 1 L, add 40 g of paraformaldehyde to heated (60 °C) 800 ml of 1x PBS. Blocking solution: 2% skim milk, 1.0% Triton X-100, 1x PBST. DMEM:Dulbecco's Modified Eagle Medium QCPN antibody (monoclonal, 1/50, Developmental Studies Hybridoma Bank HRP (horseradish peroxidase) conjugated with goat anti mouse (polyclonal, 1/200, Jackson ImmunoResearch Lab, USA DAB (3,3′-Diaminobenzidine) H2O2 (30 % hydrogen peroxide solution) AG beads (AG 1-X2 resin, 143–1255, Bio-Rad) SDF-1 protein (300-28A; PeproTech) B. Resources needed to make video Olympus dual head teaching microscope MovieZilla Movie Maker Software Samsung Galaxy S8 Smartphone