Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Thiago Cabral; Marcus A. Toral; Gabriel Velez; James E. DiCarlo; Anuradha M. Gore; MaryAnn Mahajan; Stephen H. Tsang; Alexander G. Bassuk; Vinit B. Mahajan

doi:10.3791/56203

Dissecação da Retina humana e RPE coroide para análise proteômica

JoVE Journal
Biochemistry
Author Produced

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JoVE Journal Biochemistry

Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Dissecação da Retina humana e RPE coroide para análise proteômica

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06:54 min

November 12, 2017

DOI: 10.3791/56203-v

Thiago Cabral*^1,2,7,8, Marcus A. Toral*^3,4, Gabriel Velez^3,4, James E. DiCarlo^1,2, Anuradha M. Gore³, MaryAnn Mahajan³, Stephen H. Tsang^1,2, Alexander G. Bassuk^5,6, Vinit B. Mahajan^3,9

¹Barbara & Donald Jonas Stem Cell Laboratory, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Pathology & Cell Biology, Institute of Human Nutrition, College of Physicians and Surgeons,Columbia University, ²Edward S. Harkness Eye Institute,New York-Presbyterian Hospital, ³Omics Laboratory, Byers Eye Institute, Department of Ophthalmology,Stanford University, ⁴Medical Scientist Training Program,University of Iowa, ⁵Department of Pediatrics,University of Iowa, ⁶Department of Neurology,University of Iowa, ⁷Department of Ophthalmology,Federal University of Sao Paulo (UNIFESP), ⁸Department of Ophthalmology,Federal University of EspÍrito Santo (UFES), ⁹Palo Alto Veterans Administration, Palo Alto, CA

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Overview

This article describes a method for capturing tissue biopsies from distinct regions of the human retina, including the fovea, macula, and peripheral retina. The collected samples are intended for proteomic analysis, providing insights into the molecular composition of these regions.

Key Study Components

Area of Science

Neuroscience
Proteomics
Ophthalmology

Background

The human retina has functionally and molecularly distinct regions.
Understanding these regions is crucial for various research applications.
Proteomic analysis can reveal important molecular details.
Biopsy techniques are essential for sample collection.

Purpose of Study

To develop a method for dissecting and collecting retinal regions.
To facilitate downstream proteomic analysis of retinal tissues.
To enhance understanding of retinal molecular composition.

Methods Used

Utilization of a four millimeter punch biopsy tool.
Manual removal of tissue layers from the human eye.
Biopsy collection from the fovea, macula, and peripheral retina.
Placement of the eye in a petri dish for dissection.

Main Results

Successful capture of tissue samples from targeted retinal regions.
Method demonstrated effectiveness for proteomic research.
Provided a clear protocol for future studies.
Enhanced understanding of retinal tissue composition.

Conclusions

The method allows for precise collection of retinal biopsies.
Facilitates detailed proteomic analysis of the retina.
Contributes to the understanding of retinal biology.

Frequently Asked Questions

What regions of the retina are targeted for biopsy?

The fovea, macula, and peripheral retina are targeted for biopsy.

What tool is used for the biopsy?

A four millimeter punch biopsy tool is used for the procedure.

What is the purpose of the biopsies?

The biopsies are intended for downstream proteomic analysis.

How is the human eye prepared for biopsy?

The eye is placed in a petri dish after being butterflied.

What is the significance of proteomic analysis?

Proteomic analysis helps in understanding the molecular composition of retinal tissues.

Can this method be applied to other tissues?

While this method is specific to the retina, similar techniques may be adapted for other tissues.

Is this method suitable for clinical applications?

The method is primarily designed for research purposes, but insights gained may inform clinical applications.

A retina humana é composta de regiões funcionalmente e molecularmente distintas, incluindo a fóvea, mácula e retina periférica. Aqui, descrevemos um método usando o soco de biópsias e remoção manual das camadas de tecido do olho humano para dissecar e coletar estas regiões distintas da retina para análise proteômica a jusante.

Este vídeo ilustrará como capturar biópsias de tecido da fóvea, mácula e retina periférica para pesquisa proteômica. Em várias etapas, usaremos uma ferramenta de biópsia por punção de pele de quatro milímetros. O olho humano é colocado em uma placa de Petri depois de ter sido borboleta.

A ferramenta de biópsia por punção é centralizada sobre a fóvea, pressionada para baixo, rolada levemente até que uma incisão seja feita ao redor da fóvea. Centralize e pressione uma biópsia por punção de oito milímetros dentro das arcadas da mácula com leve pressão e rolamento. Novamente, use a biópsia por punção de quatro milímetros para fazer uma série de perfurações na retina periférica do lado de fora da arcada.

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