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流式细胞仪数据分析的图形化分析,用于描述 λ 噬菌体上控制的荧光蛋白的展示。

Graphical analysis of flow cytometer data for characterizing controlled fluorescent protein display on λ phage.

机构信息

Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.

出版信息

Cytometry A. 2012 Dec;81(12):1031-9. doi: 10.1002/cyto.a.22211. Epub 2012 Oct 1.

Abstract

As native virus particles typically cannot be resolved using a flow cytometer, the general practice is to use fluorescent dyes to label the particles. In this work, an attempt was made to use a common commercial flow cytometer to characterize a phage display strategy that allows for controlled levels of protein display, in this case, eGFP. To achieve this characterization, a number of data processing steps were needed to ensure that the observed phenomena were indeed capturing differences in the phages produced. Phage display of eGFP resulted in altered side scatter and fluorescence profile, and sub-populations could be identified within what would otherwise be considered uniform populations. Surprisingly, this study has found that side scatter may be used in the future to characterize the display of nonfluorescent proteins.

摘要

由于天然病毒颗粒通常无法通过流式细胞仪进行解析,因此通常使用荧光染料对颗粒进行标记。在这项工作中,尝试使用常见的商业流式细胞仪对噬菌体展示策略进行特征描述,该策略允许控制蛋白质的展示水平,在这种情况下为 eGFP。为了实现这种表征,需要进行许多数据处理步骤,以确保观察到的现象确实可以捕捉到所产生噬菌体的差异。eGFP 的噬菌体展示导致侧向散射和荧光分布发生变化,并且可以在否则被认为是均匀群体的情况下识别亚群。令人惊讶的是,这项研究发现侧向散射将来可能用于表征非荧光蛋白的显示。

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