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定量分析与光感受器外节片段结合的 FITC 标记的乳胶珠:研究视网膜色素上皮细胞吞噬作用的简单、廉价方法。

Quantifying FITC-labeled latex beads opsonized with photoreceptor outer segment fragments: an easy and inexpensive method of investigating phagocytosis in retinal pigment epithelium cells.

机构信息

Department of Ophthalmology, University Medical Center, University of Kiel, Kiel, Germany.

出版信息

Ophthalmic Res. 2011;46(2):88-91. doi: 10.1159/000323271. Epub 2011 Jan 26.

Abstract

The retinal pigment epithelium (RPE) is the most active phagocyte in the human body, and this function is indispensable for vision. In this study, we introduce an easy and inexpensive way to quantify RPE phagocytosis by opsonizing FITC-labeled latex beads of a defined size with a crude mixture of photoreceptor outer segment fragments, using cells of porcine origin. After performance of the desired experiment, phagocytosis can easily be quantified by counting the ingested particles. With this protocol, we combine the advantage of a natural target with the advantages of the defined properties of latex beads. Moreover, in order to conduct these phagocytosis experiments, as laboratory equipment, in addition to cell culture equipment, only a centrifuge and a simple fluorescence microscope are needed.

摘要

视网膜色素上皮(RPE)是人体中最活跃的吞噬细胞,这种功能对于视觉是不可或缺的。在这项研究中,我们使用猪源细胞,通过用光感受器外节片段的粗混合物对 FITC 标记的乳胶珠进行调理,介绍了一种简单且廉价的定量 RPE 吞噬作用的方法。完成所需实验后,通过计数吞噬的颗粒,很容易定量吞噬作用。使用本方案,我们将天然靶标的优势与乳胶珠的明确定义的特性的优势相结合。此外,为了进行这些吞噬实验,除了细胞培养设备外,作为实验室设备,仅需要离心机和简单的荧光显微镜即可。

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