获得高质量的流式细胞术数据。

Q1 Health Professions Current Protocols in Cytometry Pub Date : 2020-06-01 DOI:10.1002/cpcy.74
Amy Fox, Taru S Dutt, Burton Karger, Andrés Obregón-Henao, G Brooke Anderson, Marcela Henao-Tamayo
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引用次数: 6

摘要

流式细胞术可以可视化细胞的物理、功能和/或生物学特性,包括抗原、细胞因子、大小和复杂性。随着越来越大的流式细胞仪面板能够分析多达50个参数,需要标准化流式细胞仪方案,以获得可输入分析算法的高质量数据。如果没有这些干净的数据,算法可能会错误地对样本中存在的细胞群进行分类。在本协议中,我们概述了一种综合的方法来制备用于多色流式细胞术的样品。使用多个洗涤步骤和严格的控制创建高质量的数据与细胞群之间的良好分离。使用该协议获得的实验数据可以通过执行端到端分析的计算算法进行分析。©2020 by John Wiley & Sons, Inc。基本方案1:流式细胞术单细胞悬液的制备支持方案1:肺准备支持方案2:流式细胞仪上细胞计数基本方案2:表面和细胞内流式细胞术染色支持方案3:单色珠对照。
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Acquisition of High-Quality Spectral Flow Cytometry Data.

Flow cytometry allows the visualization of physical, functional, and/or biological properties of cells including antigens, cytokines, size, and complexity. With increasingly large flow cytometry panels able to analyze up to 50 parameters, there is a need to standardize flow cytometry protocols to achieve high-quality data that can be input into analysis algorithms. Without this clean data, algorithms may incorrectly categorize the cell populations present in the samples. In this protocol, we outline a comprehensive methodology to prepare samples for polychromatic flow cytometry. The use of multiple washing steps and rigorous controls creates high-quality data with good separation between cell populations. Experimental data acquired using this protocol can be analyzed via computational algorithms that perform end-to-end analysis. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Preparation of single-cell suspension for flow cytometry Support Protocol 1: Lung preparation Support Protocol 2: Counting cells on a flow cytometer Basic Protocol 2: Surface and intracellular flow cytometry staining Support Protocol 3: Single-color bead controls.

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来源期刊
Current Protocols in Cytometry
Current Protocols in Cytometry Health Professions-Medical Laboratory Technology
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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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Issue Information Detection and Sorting of Extracellular Vesicles and Viruses Using nanoFACS Live Imaging of the Lung Small Particle Fluorescence and Light Scatter Calibration Using FCMPASS Software. Issue Information
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