Flow Cytometry — Introduction & Basics Guide
Author: Mike Blundell | Reviewer: Chloe Fenton
What Is Flow Cytometry?
Flow cytometry is a technique used to measure properties of cells in a fluid as they pass through one or more laser beams. Cells are fluorescently labeled using antibodies conjugated to fluorescent dyes, which emit light of different wavelengths upon excitation by a specific laser.
Due to its ability to rapidly identify and characterize single cells in a multiparametric manner, flow cytometry is widely used across various fields, including immunology, cancer biology, virology, and disease monitoring and diagnosis.
Flow Guide Contents
Welcome to the starting point for all flow cytometry learning at Bio-Rad. This flow cytometry guide gives you a basic overview of all the important aspects of flow cytometry. With chapters on instrumentation, useful reagents, controls, experimental set up, and much more, this guide enables best practice to be followed and gives practical advice on building multicolor panels with example protocols.
Who is it for?
Our guide is invaluable to beginners wanting to start flow cytometry and as a handy tool for teaching others about this powerful application.
To further assist learning we have recently added a Flow Cytometry glossary of terms to this guide.
Chapter 1 — Principles of the Flow Cytometer
This chapter explains how a flow cytometer works. You will learn how the cells pass through the instrument, how light is detected and measured and the basic principles behind sorting cells.
Chapter 2 — Principles of Fluorescence
Fluorophores are fluorescent markers which absorb light energy and emit at a longer wavelength. This chapter explains how they work, why fluorescent markers are so important in flow cytometry and how to compensate between them.
Chapter 3 — Data Analysis
Flow cytometry data can be analyzed in different ways. Here we give common examples of gating strategies and how data can be best represented.
Chapter 4 — Controls in Flow Cytometry
Controls are vital in any experiment to reliably distinguish your results from background variation and non-specific effects. Here we will discuss some essential controls for flow cytometry you must consider to ensure publication quality flow cytometry data.
Chapter 5 — Optimizing Your Experiments
One of the fundamentals of flow cytometry is the ability to measure single particles as they pass through the laser, however, if you start with a poor sample it is likely that the data collected will be poor as well. In this chapter we give you advice on how to prepare and treat samples to ensure you have a viable cell suspension.
Chapter 6 –— Basic Rules for Building Multicolor Panels
Building complex multicolor panels can be difficult. This chapter gives some basic rules you should follow in your experimental design to build multicolor panels.
Chapter 7 — Uses of Flow Cytometry
Flow cytometry has many varied uses. In this section we will briefly discuss popular techniques currently being used in flow cytometry to distinguish one particle from another and some of the emerging technologies.
Chapter 8 — Common Protocols
The data of quality from flow cytometry can be affected by both your cell health and staining protocol used. Here we give examples of commonly used cell preparation techniques and staining protocols for surface markers, intracellular markers and DNA.
Chapter 9 — Troubleshooting
There can be many reasons for poor results. Here we list some of the common problems with advice to help you get the best staining possible along with recommended reading if you want more information.
Flow Cytometry Basics Guide Download
Get your own copy to peruse at your leisure
Download our updated Flow Cytometry Basics Guide to have practical advice, best practice examples, and a basic overview of all the important flow cytometry principles in one handy location.
Resources
Bio-Rad is committed to helping you succeed in Flow Cytometry by sharing knowledge and best practices from our experts. Below are some extremely useful resources to enhance your journey of discovery and support your success.
All Your Flow Needs Covered
As experts in this application, we can confidently say that our Flow Cytometry Hub is the perfect place to find everything you need to achieve success with your FC experiments. From products to instrumentation, downloadables to webinars, articles to technical support, Bio-Rad has it all. Below is a small example of what our main hub section contains.