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JoVE Journal
Biochemistry
人视网膜解剖与 RPE-脉络膜的蛋白质组分析
人视网膜解剖与 RPE-脉络膜的蛋白质组分析
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

人视网膜解剖与 RPE-脉络膜的蛋白质组分析

Full Text
12,015 Views
06:54 min
November 12, 2017

DOI: 10.3791/56203-v

Thiago Cabral*1,2,7,8, Marcus A. Toral*3,4, Gabriel Velez3,4, James E. DiCarlo1,2, Anuradha M. Gore3, MaryAnn Mahajan3, Stephen H. Tsang1,2, Alexander G. Bassuk5,6, Vinit B. Mahajan3,9

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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Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

人类视网膜由功能和分子的不同区域组成, 包括黄斑、黄斑和外周视网膜。在这里, 我们描述了一个方法使用穿孔活检和手工清除组织层从人眼解剖和收集这些不同的视网膜区域的蛋白质组分析。

本视频将说明如何从中央凹、黄斑和周边视网膜捕获组织活检以进行蛋白质组学研究。在几个步骤中,我们将使用 4 毫米的皮肤穿刺活检工具。人眼在经过蝴蝶化后被放置在培养皿中。

钻取活检工具位于中央凹的正中央,向下按压,轻轻滚动,直到在中央凹周围做一个切口。在黄斑的弓内居中并按压 8 毫米的穿刺活检,轻轻按压和滚动。同样,使用 4 毫米穿刺活检在拱廊外的周边视网膜上进行一系列或穿孔。

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