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Table 1 Protocol parameters differing from the standard SpotDetector® algorithm
| Object Identification | |||
|---|---|---|---|
| Channel 1 | Method: | FixedThreshold | |
| Value: | 125 | ||
| Channel 2 | Method: | TriangThreshold | |
| Value: | 0.499 | ||
| Object Selection Parameter | Min | Max | |
| Channel 1 | ObjectArea: | 100 | 6000 |
| Channel 2 | SpotAreaCh2: | 3 | 10000 |
| SpotAvgIntenCh2: | 30 | 4095 | |
| SpotTotalIntenCh2: | 1691 | 4411431044 | |
| Assay Parameters | |||
| BackgroundCorrection: | 140 | ||
| ObjectSegmentationCh1: | 20 | ||
| SmoothFactorCh1: | 5 | ||
| SpotDetectRadiusCh2: | 15 | ||
| TargetCircModifierCh2: | 512 | ||
| PixelSize: | 0.256 microns | ||
