I've noticed that some people are using lots and lots of layers in their .psd files. It looks like the original layers they used to make thier art in the first place. psd2cssonline will do a fair job but it won't make a very optimized webpage that way. It's a good idea to merge some of those layers down to a single layer - for psd2cssonline at least. That way only a single .png file will be generated for each.
Imagine you have a complicated page with 100 layers. But theres really only 3 or 4 places where text or images or ads will go. If you are able to merge together most of those layers to (for example) a header or logo, an area for text, an area for ads and a footer - then you'll have only 4 CSS layers and 4 png files in the entire page. If you leave the psd file 100 layers, then you will have 100 CSS layers with 100 png files.
Check out the Guidelines for more info.
Thanks!
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Shaun