Table 1.
Flow cytometry controls
| Control sample | EdU | Stain | Purpose |
|---|---|---|---|
| 1. Mesonephroi × 4 |
No |
• Click-iT 647 |
Negative sample for establishing gates in the 647 nm and 488 nm channels |
| • MVH-alexa fluor 488 | |||
| • Propidium iodide | |||
| 2. Gonads × 4 |
Yes |
• Click-iT 647 |
Positive controls for setting the laser power in the 488 nm and 647 nm channels. Use females gonads for this if concentrating of male germ cell development and vice-versa if concentrating on female germ cell development (or a mix, if concentrating on both). |
| • MVH-alexa fluor 488 | |||
| • Propidium iodide | |||
| 3. Mesonephroi × 2 (optional) |
Yes |
• Click-iT 647 only |
Single color control for staining compensation |
| 4. Mesonephroi × 2 (optional) |
No |
• Propidium iodide only |
Single color control for staining compensation |
| 5. Gonads × 2 (optional) | No | • MVH-alexa fluor 488 only | Single color control for staining compensation |