从染料稀释淋巴细胞增殖实验中获取定量细胞生物学信息。
Deriving Quantitative Cell Biological Information from Dye-Dilution Lymphocyte Proliferation Experiments.
作者信息
Roy Koushik, Shokhirev Maxim Nikolaievich, Mitchell Simon, Hoffmann Alexander
机构信息
Department of Microbiology, Immunology, and Molecular Genetics, Institute for Quantitative and Computational Biosciences, University of California, Box 951570, 570 Boyer Hall, Los Angeles, CA, 90095, USA.
The Salk Institute for Biological Studies, La Jolla, CA, 92037, USA.
出版信息
Methods Mol Biol. 2018;1707:81-94. doi: 10.1007/978-1-4939-7474-0_6.
The dye-dilution assay is a powerful tool to study lymphocyte expansion dynamics. By combining time course dye-dilution experiments with computational analysis, quantitative information about cell biological parameters, such as percentage of cells dividing, time of division, and time of death, can be produced. Here, we describe the method to generate quantitative cell biological insights from dye-dilution experiments. We describe experimental methods for generating dye-dilution data with murine lymphocytes and then describe the computational data analysis workflow using a recently developed software package called FlowMax. The aim is to interpret the dye-dilution data quantitatively and objectively, such that cell biological parameters can be reported with an appropriate measure of confidence, which in turn depends on the quality and quantity of available data.
染料稀释测定法是研究淋巴细胞扩增动力学的有力工具。通过将时间进程染料稀释实验与计算分析相结合，可以得出有关细胞生物学参数的定量信息，例如细胞分裂百分比、分裂时间和死亡时间。在这里，我们描述了从染料稀释实验中获得定量细胞生物学见解的方法。我们描述了用小鼠淋巴细胞生成染料稀释数据的实验方法，然后描述了使用最近开发的名为FlowMax的软件包进行计算数据分析的工作流程。目的是对染料稀释数据进行定量和客观的解释，以便能够以适当的置信度报告细胞生物学参数，而这又取决于可用数据的质量和数量。
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