2.6 HE Staining

2.6 HE Staining

After removing the tumor tissue from the formaldehyde and rinsing it, immerse it sequentially in 70%, 80%, 95%, and 100% ethanol solutions for 3 hours, 2 hours, 1.5 hours, and 1.5 hours respectively, to dehydrate it; then immerse it in xylene for 2 hours, remove it and soak it in fresh xylene solution for 1 hour, followed by soaking in a xylene: soft wax solution (1:1) for 2 hours, and finally embed the tumor in wax until it solidifies. After the wax blocks are prepared, use a microtome to cut slices and place them on anti-detachment slides.

Research on Pei Zhengxue’s series of prescriptions

Subsequently, the slices are subjected to HE staining, with the following steps: A. Immerse the slices sequentially in 3 cups of xylene for 10 minutes each, then in 100%, 100%, 95%, 80%, and 70% ethanol solutions for 2 minutes each, finally placing them in distilled water for 2 minutes to remove the wax → B. Place the slices in hematoxylin for 15 minutes to stain, then rinse them → C. Immerse the slices again in 1% hydrochloric acid alcohol for 15 seconds to differentiate the colors → D. Rinse them under running water for 5 minutes to reverse the blue color → E. Re-stain with eosin for 13 minutes, then rinse again → F. Immerse the slices again in 70% and 80% ethanol for 30 seconds each, in 90% and 95% ethanol for 1 minute each, and in 100% ethanol for 2 minutes each to remove moisture from the tissue → G. Make the tissue transparent by immersing the slices in an alcohol:xylene (1:1) solution and in xylene III for 2 minutes each → H. Finally, mount the slides and observe the changes in tumor cell morphology under the microscope.