3 – Data analysis

6 Flow Cytometry Gating Tips That Most Scientists Forget

By: Tim Bushnell, PhD

After completing the perfect staining and cytometry run, the hard work begins – data analysis.  To properly identify the cells of interest, it is critical to pull together knowledge of the biology with the controls run in the experiment to properly place the regions of interest that will be dictate the final results.  Gating is an all-or-nothing data reduction process.  Cells inside the gate move to the next checkpoint, while cells outside the gate – even by a pixel, are excluded. 1.  Before beginning, know as much as you can about the populations of interest. While it may sound flip,…

5 Important Tips For Analyzing Your Data

By: Tim Bushnell, PhD

Depending on the experimental design, many researchers will be doing complex assays that will require statistical analysis to determine if the hypothesis being tested is statistically significant or not. Unfortunately, many researchers go about this analysis the wrong way, resulting in spurious conclusions. The following points are guides to help think about the steps necessary in flow cytometry data analysis. 1. Before you start Define your hypothesis. This may sound simplistic, but understanding the purpose of the experiments is the first step in performing good statistical analysis. Stating the hypothesis will allow the researcher to choose the correct statistical test…

How To Create Flow Cytometry Gates

By: Tim Bushnell, PhD

After completing the perfect staining and cytometry run, the hard work begins – data analysis. To properly identify the cells of interest, it is critical to pull together knowledge of the biology with the controls run in the experiment to properly place the regions of interest that will be dictate the final results. Gating is an all-or-nothing data reduction process. Cells inside the gate move to the next checkpoint, while cells outside the gate – even by a pixel, are excluded. 1. Before beginning, know the populations of interest. While it may sound flip, knowing what cells are the target…

Flow Cytometry Statistics

By: Tim Bushnell, PhD

Understanding statistics and fow cytometry statistical analysis is critical to understanding flow cytometry data. One of the powers of flow cytometry is the fact that we generate large amounts of data that are amenable to statistical analysis of our populations of interest.  Using the standard set of statistical analysis tools allows for hypothesis testing and ultimately determining if there is statistical significance in the datasets. There are two basic classes of questions that are typically asked in flow cytometry.  The first class relate to changes in the number or percent of a specific population upon treatment or disease state.  A…

FACS Analysis

By: Tim Bushnell, PhD

Flow cytometry is the science of measure the physical and biochemical processes on cells and cell-like particles. This analysis is performed in an instrument called the flow cytometer.  FACS Analysis is the short-hand expression for this type of cell analysis The term FACS stands for Fluorescent Activated Cell Sorting, a term first coined by Len Herzenberg in the 1970’s, and later trademarked by Becton Dickinson. Since that time, FACS has come to be used as a generic term for all of flow cytometry, even though it is a specific trademarked term.