Cytometry and Antibody Technology

Training courses

Flow Basics

Download the presentation

All users who want to use instruments unassisted must complete the required Flow Basics training. The training is in two parts: a 1hr course called Flow Basics and a one-on-one training session on a cytometer.

The Flow Basics Class is presented every other Wednesday
Time: 10:00AM to 11:30AM.
Next classes: May 5th, May 19th

Register here

You will receive your Zoom invitation at 11:30AM the day before the presentation.

Contact the CAT Facility to schedule your hands-on training.

 

 

Software Tutorial

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.

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

Spectral Flow Cytometry

Users who want to use the spectral flow cytometer (Cytek Aurora) are required to go through additional training with Laura Johnston after completing the original Flow Basics course. These videos may be used as a reference, but are not a replacement for the required training. More information on spectral flow cytometry can be found here.

An abbreviated training video is also available for Principal Investigators or other researchers who will be analyzing spectral flow cytometery data but not running the instrument.

Spectral Training Course

View playlist on YouTube

Download the slides (may be different than the videos)

Spectral Training/Troubleshooting for Principal Investigators