How to perform a 'Punctate Count'

Area mode:

Step 1: When your chosen image has been imported into the viewing area, select 'Staining', followed by 'Area' and then 'Detect staining', Figure 1.

Step 2: This will activate the drawing tool. Using the mouse place the cursor on one corner of the area of interest and left click (indicated by a small square), Figure 1.

Figure 1

Step 3: From here, continue to move the mouse and left click in intervals to form a border around the desired area, Figure 1.

This mode only creates straight lines, therefore, when covering more detailed areas additional anchor points will be required to improve accuracy.

Step 4: To finish, make your way around to the first anchor point and left click to complete the selection.  The first anchor point will be visible as a large square, Figure 1. 

Figure 2

Step 5: When the area is closed, the area highlights the punctates in red, Figure 2. The results are displayed in the results panel, Figure 3. 

Figure 3

 Corneal Grid mode:

Step 1: Alternatively, on the same image you have chosen, select the 'Corneal Grid' button followed by 'Detect Staining'.

Step 2: Left click on the screen and a 5 zones corneal grid will appear in the viewing area, Figure 4. 

Altering the Corneal Grid 

Step 1: To position the grid in the desired area, move the cursor into the inner circle, then click and drag.

Step 2: Once in position, it will automatically detect the punctates, which can be viewed in the results panel. Each result displayed relates to one of the five zones illustrated in figure 4.

Step 3: On the viewing area, you will see the staining highlighted in red, Figure 4. This provides a visual representation of the collected data in the results panel.

Figure 4

 How to remove artifacts on the image displayed: 

Step 1: Click the 'Mark Artifact' icon as shown in Figure 5 below. 

Step 2: Select the artifact(s) you would like to erase by clicking on the red dots. On completion the selected dots will appear blue, Figure 5.

Step 3: Once you have selected the artifact(s) you wish to delete, just click the 'Erase Artifact', Figure 6. 

Figure 5

Figure 6