Preparation for microdissection (Paraffin/Frozen)

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom

Basic protocol for Eosin & Haematoxylin histological preparation of microdissection slide.

• For paraffin-embedded sections, place slides into fresh xylene to remove parrafin.
• Place slide into fresh xyline
• Transfer slide to 100% ethanol, 95% ethanol, then 70% ethanol; 30 seconds/step
• Transfer slide into de-ionized water
• Transfer slide into Mayer’s Haematoxylin
• Return to de-ionized water to rinse for 15 seconds x2 changing dH20 between steps
• Transfer slide to 70% ethanol
• Transfer slide into Eosin Y
• Return to de-ionized water to rinse for 30 seconds x2
• To reduce brittleness of tissue, place in 3% glycerol in de-ionized water
• Shake and air dry to remove layer of glycerol and water
• Microdissection is best performed in the following 5-10 minutes following preparation. If if tissue dries return tissue to 3% glycerol/water; step #10