To successfully run the assay, perform the following steps:
Click on the button from the home page. The Assay tab opens.
Select an enabled assay. Information about the assay and used dyes is shown.
Adjust the Assay Settings in the right. Click .
Insert the well plate into the microscope stage and fill in information about it (Name and Plate are obligatory).
Click . The system automatically detects the well plate, locates the first well containing cells, performs a basic auto focus on it, and then scans the entire well plate using brightfield optics. An overview of all wells is captured, and the system automatically selects the wells that contain cells.
Click and select wells or confirm the current selection on which wells the Autosignal.ai will run. Confirm the selection by clicking .
Note
Do this step only when Preview Type: User defined Wells was selected.
Click and select the wells for capturing (Scanning phase). Confirm the selection by clicking .
Click click into the well plate to add a label selected on the left. More labels can be added by clicking
. Color/name can be changed by clicking on the label color/name itself. Delete the label by clicking 
next to it.Switch to Dosing and insert dosage numbers directly into the selected well. The dosing selection can be expanded by dragging the yellow line to automatically fill in the neighboring wells.
To save the label/dosing definition into the database, click
in the top right corner and use the saving manager.Confirm the labels and dosing by clicking .
Click . Autosignal.ai is executed and all channels required for the assay are captured.
If Run analysis was checked in the Assay settings, the analysis is performed on the captured data and the results are shown both for the whole well plate and for the single (selected) well. Results can be inspected and exported using the automatic report.
Note
Assay has to be enabled by the administrator before it can be used in the Run Assay window.
Click on the button and the Assay Definition window opens.
Select an assay and optionally adjust its parameters.
Click . Assay is now enabled for the user.
Assay Settings
If turned on, an e-mail notification is sent to each user in the group during and after the Scanning phase.
If turned on, analysis is executed immediately after the Scanning phase.
User defined wells - Autosignal.ai finds the best brightness value for the well plate only on wells selected by the user.
All wells selected for acquisition - performs Autosignal.ai on all wells to obtain the most optimal brightness value.
Last settings - the last option selected.
If Auto. continue after preview step is turned on, the application executes the Scanning phase directly after the Preview phase.
Scan Area Selection defines which wells are selected for the scanning phase.
If Use Anomaly detection is set to YES, Morphology and/or Condensation options may appear depending on the assay type. Morphology uses the Nikon AI (artificial intelligence) module to evaluate whether the cell shape is normal or anomalous and labels the wells “Morphology Anomaly”. Condensation uses Nikon AI module to detect condensation on the well bottom of each well in the well plate and labels the wells “Condensation Anomaly”. Both detections are run only on the wells selected in the Acquisition Selection using the Brightfield image.