使用 FCMPASS 软件进行小颗粒荧光和光散射校准。

Q1 Health Professions
Joshua A Welsh, Jennifer C Jones
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引用次数: 0

摘要

使用流式细胞仪分析小颗粒已有几十年的历史。最近,由于传统和市售流式细胞仪的灵敏度提高,加上人们对细胞外囊泡 (EV) 等小颗粒的兴趣与日俱增,小颗粒分析的应用越来越广泛。尽管小颗粒流式细胞仪的应用越来越广泛,但由于数据报告缺乏标准化,导致有关 EVs 的文献越来越多,而这些文献却不容易解释、验证或复制。荧光和光散射标准化的方法已经非常成熟,进行这些分析的试剂也可以在市场上买到。在此,我们介绍 FCMPASS 软件包,它可对小颗粒进行荧光和光散射校准,同时生成符合 MIFlowCyt-EV 标准报告框架的标准报告。本文介绍了使用 FCMPASS 实施校准的工作流程如下:获取荧光和光散射校准材料、为软件中使用的参考材料编目、创建细胞仪数据库和数据集以关联校准数据和 fcs 文件、导入 fcs 文件进行校准、输入荧光校准参数、输入光散射校准参数以及将校准应用于 fcs 文件。2020 年出版。美国政府。基本规程 1:光散射校准材料的获取和选通 基本规程 2:荧光校准材料的获取和选通 替代规程:基本规程 3:光散射校准材料编目 基本规程 4:荧光校准材料编目 基本规程 5:创建细胞仪数据库和数据集 基本规程 6:导入 fcs 文件 基本规程 7:荧光校准 基本规程 8:光散射校准 基本规程 9:执行和报告 fcs 文件校准。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Small Particle Fluorescence and Light Scatter Calibration Using FCMPASS Software.

Use of flow cytometry to analyze small particles has been implemented for several decades. More recently, small particle analysis has become increasingly utilized owing to the increased sensitivity of conventional and commercially available flow cytometers along with growing interest in small particles such as extracellular vesicles (EVs). Despite an increase in small particle flow cytometry utilization, a lack of standardization in data reporting has resulted in a growing body of literature regarding EVs that cannot be easily interpreted, validated, or reproduced. Methods for fluorescence and light scatter standardization are well established, and the reagents to perform these analyses are commercially available. Here, we describe FCMPASS , a software package for performing fluorescence and light scatter calibration of small particles while generating standard reports conforming to the MIFlowCyt-EV standard reporting framework. This article covers the workflow of implementing calibration using FCMPASS as follows: acquisition of fluorescence and light scatter calibration materials, cataloguing the reference materials for use in the software, creating cytometer databases and datasets to associate calibration data and fcs files, importing fcs files for calibration, inputting fluorescence calibration parameters, inputting light scatter calibration parameters, and applying the calibration to fcs files. Published 2020. U.S. Government. Basic Protocol 1: Acquisition and gating of light scatter calibration materials Basic Protocol 2: Acquisition and gating of fluorescence calibration materials Alternate Protocol: Cross-calibration of fluorescence reference materials Basic Protocol 3: Cataloguing light scatter calibration materials Basic Protocol 4: Cataloguing fluorescence calibration materials Basic Protocol 5: Creating cytometer databases and datasets Basic Protocol 6: Importing fcs files Basic Protocol 7: Fluorescence calibration Basic Protocol 8: Light scatter calibration Basic Protocol 9: Performing and reporting fcs file calibration.

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来源期刊
Current Protocols in Cytometry
Current Protocols in Cytometry Health Professions-Medical Laboratory Technology
自引率
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期刊介绍: Published in affiliation with the International Society for Advancement of Cytometry, Current Protocols in Cytometry is a "best practices" collection that distills and organizes the absolute latest techniques from the top cytometry labs and specialists worldwide. It is the most complete set of peer-reviewed protocols for flow and image cytometry available.
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