TABLE 2.

Troubleshooting table.

Step	Problem	Possible reason	Solution
18	50 μl of medium does not stay on coverslips	The coverslips do not recover to room temperature and are still wet	Leave the plates at room temperature longer or leave the lid half open for quick evaporation
19	The coverslips dry out	The drop of medium has drained off the coverslip	When moving and transferring the plate, make sure this is done gently. Ensure that the incubator door is opened and closed gently
22	No PDGFRα stained	The cells are overfixed or not fixed properly	Stain for the membrane protein before fixation as described in the notes of Table 1
23	No 04+ cells	04 cells died and detached; or inappropriate staining procedure	Adding 0.5% (vol/vol) fetal bovine serum into the culture medium may help the survival of 04+ cells. Incubate the cells with 04 antibody for 15 min before fixing the cells with 4% PFA (wt/vol)

PDGFRα, platelet-derived growth factor receptor alpha; PFA, paraformaldehyde.