Table 1.
Function of the methods within the find region building blocks on the Opera and Operetta imaging systems contained within the amastigote assay script. The parameters used within the developed script to identify parasites within host cells are shown for each building block of nuclear, cytoplasm and spot analyses.
| Find region (method) | Method | Function | Parameters available | Parameter used |
|---|---|---|---|---|
| Nuclei (C) | Roundness | Estimates the roundness of the detected nuclei | −2 to 3 | 0.6 |
| Common threshold | The lower level of pixel intensity for the whole image that may belong to nuclei | 0–1 | 0 (no constriction) | |
| Area | Estimates the area of the detected nuclei | −100 to 200 | 80 | |
| Cytoplasm (A) | Common threshold | Parameter determining first guess borders of the cytoplasm | 0–1 | 0 (no constriction) |
| Cytoplasm (A) | Select cell region | Tuning the inner border, from nuclear to cell border | −200 to 100 | 50 |
| Spot detection (C) | Radius | Determines the radius of radius of spot-like features | 0–55 | 2.2 |
| Contrast | Determines how intense a spot must be to be detected | 0–1 | 0.13 | |
| Spot to region intensity | Sets a lower threshold for spot to cell intensity. The ratio of the spot peak intensity to the mean intensity of the cell region that is searched. | 0–10 | 1.6 |