Periodic acid Schiff hematoxylin (PASH) staining protocol is adapted from other tissues to identify carbohydrate in human retinal sections in our lab. This method is used for labeling 10-12 µm thick cryosections of human retina, RPE, choroid, and sclera, preserved within 6 hours of death in 4% paraformaldehyde in 0.1 M phosphate buffer, picked up on SuperFrost glass slides, and stored at -80°C. It is intended to be counterstained for nucleic acids in nuclei using modified Harris hematoxylin.
The most common application is for demonstrating glycogen in liver tissue. Positive staining for glycogen is magenta, and nuclei stained by hematoxylin are blue.
This PASH staining protocol is helpful to identify drusen, basal laminar deposits, retinal pigment epithelium (RPE, containing abundant lipofuscin), nuclei, and cell layers of the retina, choroid, and sclera. It is preferable to traditional H&E staining for structures and pathology specific to age related macular degeneration (AMD). It is particularly useful for diagnosis and comparison to clinical retina imaging, especially optical coherence tomography. Since the late 1960s, PASH applied to human AMD specimens was noted to stain an eosinophilic material at the base of the RPE (now known as basal laminar deposit), drusen (the characteristic lesions), and Bruch’s membrane. [1-4]
1.Gass JDM. Pathogenesis of disciform detachment of the neuroepithelium. III. Senile disciform macular degeneration. Am. J. Ophthalmol. 1967;63:617-644. PMID 6019308
2.Farkas TG, Sylvester V, Archer D, Altona M. The histochemistry of drusen. American Journal of Ophthalmology. 1971;71(6):1206-1215. PMID 4253686
3.Sarks SH. Ageing and degeneration in the macular region: a clinico-pathological study. Br. J. Ophthalmol. 1976;60(5):324-341. PMID 952802
4.Vogt SD, Curcio CA, Wang L, Li C-M, McGwin G, Jr, Medeiros NE, Philp NJ, Kimble JA, Read RW. Retinal pigment epithelial expression of complement regulator CD46 is altered early in the course of geographic atrophy. Exp Eye Res. 2011;93( 4):413-423 PMID 21684273