Hi Daniel,
I took a look at your sample application you sent to support and I have the following observations:
1. Your image is located on the root folder of your application. Visual WebGui has default folders for Icons and Images that are Resources/Images and Resources/Icons. The default folders can be seen in web.config. I created a Resources folder and in there I created a Icons folder and placed the image there.
2. The TreeNode property does have an Image property, but if you set it in designer it will not be saved. In version 6.4 I checked, that this property has been removed from the designer. You can also set it by code on individual TreeNodes if you wish, but it does not appear in Intellisense. Instead, you should add an ImageList to your form, add your images to it and then select image from there to your nodes, if you need to do this in designer.
So, by moving your image, according to 1 above, and adding an imagelist + assigning images from there to your TreeNodes, according to 2 your app works fine. Tested in 6.3.8a and 6.3.10.
Please confirm if these changes make it work for you.
Palli