Post-IMS H&E Staining for Pancreas or Eye Cryosections

Carrie Romer, Jamie Allen, Jeff Spraggins, Danielle Gutierrez, Diane Saunders, Angela R.S. Kruse, Alvin Powers

Published: 2022-05-25 DOI: 10.17504/protocols.io.x54v9yw4pg3e/v1

HuBMAP

BIOMIC

Vanderbilt

Pancreas

H&E staining

Eye

Retina

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Abstract

This protocol for H&E staining can be applied to either fixed or unfixed frozen cryosections.

Steps

Matrix Removal

1.

Collect post-IMS autofluorescence

2.

Remove matrix from tissue sections by incubating in 100% alcohol, dipping gently for 0h 0m 30s. Repeat this step 2-3 times until there is no visible matrix on the slides.

H&E Staining

3.

Incubate in 95% alcohol for 0h 1m 0s , then wash, dipping until clear.

4.

Incubate in hematoxylin for 0h 0m 30s , then wash until clear.

5.

Dip 3-4x in bluing solution , then wash for 0h 0m 30s

6.

Dip 3-4x in 95% alcohol.

7.

Dip 1-2x in eosin.

8.

Dip 5-10x in 95% alcohol , then repeat using 2nd aliquot.

9.

Dip 5-10x in 100% alcohol , then repeat using 2nd aliquot.

10.

Dip 5-10x in xylene , then repeat using 2nd aliquot.

11.

Apply coverslip and seal.

Figure 1. H&E stained pancreas with magnified section showing nuclei in blue and cytoplasm in pink.
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.
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.

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