This protocol for H&E staining can be applied to either fixed or unfixed frozen cryosections.
Steps
1.
Air dry sections for 0h 5m 0s .
2.
Incubate in 95% alcohol for 0h 1m 0s , then wash, dipping until clear.
3.
Incubate in hematoxylin for 0h 0m 30s , then wash until clear.
4.
Dip 3-4x in bluing solution , then wash for 0h 0m 30s
5.
Dip 3-4x in 95% alcohol.
6.
Dip 1-2x in eosin.
7.
Dip 5-10x in 95% alcohol , then repeat using 2nd aliquot.
8.
Dip 5-10x in 100% alcohol , then repeat using 2nd aliquot.
9.
Dip 5-10x in xylene , then repeat using 2nd aliquot.
10.
Apply coverslip and seal.
Figure 1. H&E stained pancreas with magnified section showing nuclei in blue and cytoplasm in pink.Figure 2. H&E stain of human retina tissue from a 72 year old donor. Dark purple nuclear staining and pink cytoplasmic staining allow the user to readily distinguish retinal layers such as (from bottom to top) the: sclera, choroid, retinal pigmented epithelium, photoreceptor layer, outer nuclear layer, outer plexiform layer, inner nuclear layer, inner plexiform layer, ganglion cell layer, and nerve fiber layer.
