Cutting thicker sections where the stain comes out perfect every time.
Thicker pieces need extra time to dehydrate. This means more time in the first alcohol is a must. Normal time in the first alcohol is 30-60 seconds, increase this time to 90-120 seconds to draw all the water out of the dense tissue.
Add time to your hematoxylin. Doubling the time should help the stain penetrate the thick tissue. Be sure to use extra washing dips in the waters following the hematoxylin. It is important to perform these extra dips to insure the hematoxylin is properly blued.
Your Eosin will also need to be increased, add about 50% of what is used normally. For example, if you were using one dip, use one plus a quick second dip. If two dips was your norm add one extra dip for a thick cut.
The last set of alcohols need extra dips as well, about double what you use for tissue of normal thickness.
Your slides should look much better under the microscope after following these steps.
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