Interpreting Flow Cytometry Data 101 – How To Achieve Cytometry Analysis

Introduction to Interpreting Flow Cytometry Data

In flow cytometry, every cell is identified as a singular event through light emissions.

Properties of that cell can be identified through analysing the direction and activity of the light wavelengths that have passed through.

The machines that emit such lasers into cell samples churn out an inordinate amount of data and numbers which need to be run through a computer to make sense of.

Generally, all flow cytometry analysis software programmes will display this data on histograms and dot-plots for you.

 

Applications of Flow Cytometry Analysis

There are numerous applications for flow cytometry in different fields of science and research.

Immunologists use flow cytometry to analyse immune cells by size and morphology.

Immunophenotyping uses blood, bone marrow, or lymph cells to diagnose haematological malignancies.

Cell cycle analysis can help identify abnormal cell aneuploidy which can be linked to chromosomal anomalies.

But how does it work?

 

Light Scatter in Cytometry Analysis

Different directions of light scatter in the flow cytometer relate to different purposes of analysis.

Forward scatter detects the size of cells due to the change in the light path when the cell is in front.

Side scatter tells you about the granularity and complexity of the sample.

The data these light methods provide is then collected.

 

Flow Cytometry Analysis

There are benefits to both histograms and dot-plots in flow cytometry analysis.

Histograms are best when you are only looking at a single parameter, whereas dot-plots can be two or three dimensional and are best for looking at multiple parameters.

You can of course use both of these, and analysis is of course not limited to this.

Gating allows for the analysis of almost an infinite number of parameters, and allows a much more detailed view of selected cells.

Compensation is also useful in reducing the effect of spillover between parameters to only show signals once and prevent the confusion of data.

 

VenturiOne – Flow Cytometry Analysis

Our FACS Data Analysis Software, VenturiOne, provides easy an informative analysis of the largest FCS data files.

You can interpret your data in a range of ways, with the ability to generate 2D and 3D overlay plots with custom fields, statistical calculations, and rules, as well as our compensation wizard, hyperlog sliders, and gating.

Sign up for a free 30 day trial and see for yourself.

 

 

 

 

 

 

 

 

spectral flow cytometry