Staining: Thickness
How to use the 'Thickness' function
'Staining Thickness' can help visualize the depth of fluorescein over the eye by creating a heat map. Cold colours (blue) refer to thin levels of fluorescein, while warm colours (orange and red) indicate the fluorescein getting progressively thicker.
2D Analysis: Whole Image
Step 1: When your chosen image has been imported into the viewing area, select 'Staining', followed by 'Thickness'.
Step 2: To get a visual representation of the fluorescein thickness simply click '2D', Figure 1.

Figure 1
2D Analysis: Area of Interest
Step 1: Alternatively, you can draw an area of interest and generate the heat map on the selected area.
Step 2: When your chosen image has been imported into the viewing area, select 'Staining', followed by 'Thickness'.
Step 3: Using the mouse, start from one corner of the area of interest and left click to place an anchor point (indicated by a small square).
Step 4: From here continue to move the mouse and left click in intervals to form a border around the desired area.
Step 5: 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.
Step 6: When the area is closed, the heatmap is applied only to the area drawn, Figure 2.
Note: This mode only creates straight lines, therefore, when covering more detailed areas additional anchor points will be required to improve accuracy.

Figure 2
3D Analysis: Whole Image
When your chosen image has been imported into the viewing area, select 'Staining', followed by 'Thickness'. To associate the heat map colours to the thickness of fluorescein, click on the 3D button, Figure 3.

Figure 3
3D Analysis: Area of Interest
It is also possible to recreate a 3D view of the fluorescein thickness and the heat map, the red, higher ‘mountains’ representing the higher levels of fluorescein, as shown in Figure 4.
Step 1: Follow steps 1-5 outlined in ‘2D Analysis: Area of Interest’ above.
Step 2: Click 3D button, Figure 4.

Figure 4