| Experiment | ER depletion using a Red-CES2 | Stimulation of neurons using a CES2 construct | High spatial resolution imaging, CES2 |
| Objective(N.A) | 20x water (0.7) | 20x water (0.7) | 40x oil/63x oil (≥1,3) |
| 473 nm Laser | |||
| Power | 1% | 1% | |
| HVA | 600-780 | 600-780 | 400-780 |
| GainB | 0-1% | 1-2% | 0% |
| Offset | 0 | 0 | 0 |
| 559 nm Laser | |||
| power | 1% | optionalC | optionalC |
| HVA | 600-780 | optionalC | optionalC |
| GainB | 0-1% | optionalC | optionalC |
| Offset | 0 | optionalC | optionalC |
| Scanning speed | 1-2Hz | Free run | Free run |
| Picture size | 320x320 pixel | 240x240 pixel | 512x512 pixel |
| Scanning type | In line | Bidirectional | Nyquistcriterion (2 pixels/element) |
| Stimulation | 100-200 μM ATP:5-15 sec; 200-500 μM glutamate for 5-15 sec | 100-200 μM Agonist: 5-15 sec | |
| Pinhole | 2-5 airy discs | 2-5 airy discs | 1 airy disc |
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