The goal of this titration experiment is to determine the concentration of the anti-CD8FITC reagent that achieves saturating staining i.e., maximal staining of the CD8 T cells This data can be found on the FlowJo CD in the Basic Tutorial folder or you can The data files were aquired with 10,000 events collected for each sample. Cells were stained for 20 minutes, washed and resuspended in Seven tubes were stained with a serial two-fold dilution of the anti-CD8-FITC reagent Human peripheral blood mononuclear cell preparation from a single healthy donor A titration experiment wasĬarried out to determine the appropriate reagent concentration for sample staining. This reagent was produced by conjugating purifiedĪnti-CD8 antibody to the fluorochrome Flourescein (FITC). The set of data files used to illustrate this tutorial consists of an experiment titrating a Workspace files can be found in the “Basic Tutorial” folder on the FlowJo CD or at Follow along with the provided data files to get aįeel of how FlowJo works! If you wish to skip steps, we provide premade Workspaceįiles that correspond to each analysis step. Instructions to guide you through the analysis of a basic immunophenotyping experiment are contained in this tutorial. Reports that summarize multiple data samples with a click of a button. FlowJo also produces tables of statistics and graphical The repetitive tasks required to analyze flow data are greatly simplified by FlowJo.īatch processing is used to apply analyses (gated subpopulations and statistics) to Being able to view the summary of an entire experiment in a Workspace window means that there is no need to keep track of gates or statistics and your data analysis is organized in a biologically meaningful way. Nodes are organized in a hierarchical manner so that it is easy to distinguish the relationship between each population. The Workspace window not only lists all the sample files being analyzed, butĪlso displays each gated subpopulation and statistic as a node under the sample name. Handles data analysis are a direct result of these differences.įlowJo organizes and displays all the data files from an experiment in a Workspace The power of FlowJo and the ease with which it Learning to use FlowJo to efficiently analyze your flow experiments is not difficult,Īlthough it is worth noting that there are several differences between FlowJo and otherįlow cytometry analysis packages. Generate a table of statistics from all samples. Generate a graphical report including all samples.Ħ. Copy gates and statistics to all samples.ĥ. Analyze the control sample in detail (gate subsets and add statistics).ģ. Involved in analyzing a basic immunophenotyping experiment.Ģ. This tutorial is designed to introduce you to FlowJo and to the 6 steps You can use FlowJo toĪnalyze all of your flow cytometry data - regardless of the cytometer used to collect 1997-2002.įlowJo software is used for the analysis of flow cytometry data. 1997-2002.įlowJo Tutorial and Web Site are Copyright © Tree Star, Inc. University, 1996-97įlowJo is Copyright © Tree Star, Inc. We are indebted to the Herzenberg laboratory at Stanford and our active andĮnthusiastic users for their ideas, discussions, and tireless testing of new versions.įlowJo is Copyright © Trustees of Leland Stanford Jr. įlowJo Basic Tutorial by Jennifer Wilshire.įlowJo was written by Adam Treister and Mario Roederer, based on conceptsĭeveloped with David Parks, Martin Bigos, and Wayne Moore. Step 3: Copy gates and statistics to all samples. Step 2: Analyze the control sample in detail. A FITC anti-CD8 reagent was titrated to determine the Is designed to introduce you to FlowJo by demonstrating the analysis of a FlowJo software is used for the analysis of flow cytometry data.
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