Cytometry and Antibody Technology

Flow Basics 2.0

Supplementary Training Course

Home » Resources Library » Flow Basics 2.0

Flow Basics 2.0 is a series of courses that builds on the original Flow Basics course. This series outlines all of the practical steps for setting up a flow cytometry experiment and highlights factors that impact final results. The videos can be watched on our YouTube channel and the slides can be downloaded below.

There is also a good series of blog posts called “Avoid Bad Data” and a list of Best Flow Cytometry Publications in 2022.

Flow Basics 2.1: Staining Protocol and Reproducibility

  • Overview of a basic cell surface marker staining protocol
  • How does Fc block work
  • Five factors that affect antibody staining
  • What is the difference between the available viability dyes
  • Fixation
  • Download Flow Basics 2.1

Flow Basics 2.2: Optimizing the Staining Protocol

  • Why you should optimize tissue digestion
  • Alternative blocking options
  • How to determine the number of cells to stain for an experiment
  • Why and how to perform antibody titration
  • Download Flow Basics 2.2

Flow Basics 2.3: Panel Design

  • Information needed before beginning panel design
  • Tools available for for panel design
  • How to assign markers to fluorophores
  • Which fluorophores to avoid
  • Strategies for placing high spillover fluorophores
  • Download Flow basics 2.3

Flow Basics 2.4: Experiment Design

Flow Basics 2.5: Instrument setup and Automated Compensation

  • Why organizing the experimental layout is useful
  • Shortcuts for labeling tubes quickly in BD FACSDiva
  • How to set voltages on the cytometer
  • How to properly use automated compensation tools and avoid compensation errors
  • How to identify compensation errors
  • Download Flow Basics 2.5