IMR Press / FBE / Volume 4 / Issue 4 / DOI: 10.2741/e481

Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with Frontiers in Bioscience.

Open Access Article

A modified histoimmunochemistry-assisted method for in situ RPE evaluation

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1 Department of Ophthalmology, Shanghai Tenth People’s Hospital, and Tongji Eye Institute, Tongji University School of Medicine, Shanghai, China
2 Department of Ophthalmology, Second Affiliated Hospital of Soochow University, Suzhou, China
3 Department of Ophthalmology, Drexel University College of Medicine, Philadelphia, PA, USA
4 Stem Cell Research Center and Department of Regenerative Medicine, Tongji University School of Medicine, Shanghai, China

*Author to whom correspondence should be addressed.

Academic Editor: Qingli Li

Front. Biosci. (Elite Ed) 2012, 4(4), 1571–1581; https://doi.org/10.2741/e481
Published: 1 January 2012
(This article belongs to the Special Issue Hyperspectral imaging in biomedicine)
Abstract

We have developed a method for morphological evaluation of retinal pigment epithelium (RPE) with minimal perturbation of RPE-Bruch’s membranechoriocapillaris complex (RBCC). To prepare RBCC, the anterior segments of the eye were removed. The remaining eye cup was radially cut into 4 to 6 pieces from periphery to the optic nerve head. Each piece was carefully dissected into 3 parts: sclera, RBCC and neurosensory retina. RBCC flatmount with RPE monolayer facing up was formed by several relaxing radial cuts. After immuno-staining with tissue specific markers, the RBCC could be distinguished as the superficial RPE monolayer and the underneath choriocapillaris layers by fluorescence microscopy. The density, distribution, and morphology of RPE cells varied among species. This method may have brought several advantages for RPE screening over other means, because of its straight forward approach, minimal manipulation of samples, plus there is no requirement for bleaching, which result in high efficiency for result readout. For a complete morphological study of RPE in situ, this method may be combined with other methods, such as cryosections, scanning electron microscopy, etc.

Keywords
Choriocapillaris
CCE
Retinal pigment epithelium
RPE
RPE-Bruch’s membrane-choriocapillaris complex
RBCC
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