Nanocourse: Basics of flow cytometry

The FACS Facility is offering a 2 day in-house course on "Basics of flow cytometry". The course is designed to provide a comprehensive understanding of the core principles of flow cytometry, while also emphasizing practical considerations related to sample preparation and the most frequently used assays. The course is intended for staff scientists, TAs, PhD students, masters students and postdocs who want to gain the knowledge and skills necessary to effectively design, execute and interpret flow cytometry experiments.

When? Tuesday, September 2nd from 10 AM - 5 PM and Wednesday, September 3rd from 10 AM - 3 PM

Where? Seminar Room 4

Seats are limited, please sign up until Tuesday, 5th of August using the following link:

https://nc.molgen.mpg.de/cloud/index.php/apps/forms/JpowcXxqijjmmr8D

The FACS team is looking forward to seeing you there. If you have any questions regarding the course content or if it's suitable for you, please don't hesitate to contact them at facs@molgen.mpg.de.

Programme

Day 1: Introduction and core pinciples

Morning Session

Welcome and Objectives

• Brief overview of the course structure and goals.
• History of flow cytometry

• Overview of a flow cytometer and its components
  
• Key concepts: basics of fluorescence and fluorochromes
• Fluidics and optics
• PMT
• Know your cytometer: Aria II vs Aria Fusion
• Quality control
• Droplet sort theory

Afternoon Session

Experimental design

• Hypothesis/ needs
• Target cells
• Controls
• Gating strategy

Sample preparation

• Obtaining a single cell suspension, buffers, washes, filtering, temperature, antibody titration, storage and transportation
• The evilness of dead cells and introduction of different viability dyes
• Fixation
• Permeabilisation
• Downstream analysis is important to consider: how to prep for mass spec or RNAseq

Troubleshooting

• Overview of some frequently encountered problems and how to solve them (no events appearing, cells forming clumps, clogs, non-specific stainings…)

Day 2: Applications and future of flow cytometry, in the MPI and outside

Morning Session

Applications

• Phenotyping
• Apoptosis
• Proliferation
• Cell cycle
• Quantification
• Enrichment

Future of flow cytometry

• Introduction to spectral flow cytometry, differences with conventional flow cytometry
• Imaging flow cytometry – a brief overview of the S8
• Label-free flow cytometers, AI based sorting

Afternoon Session

Data analysis teaser

• Introduction to FlowJo

Wrap-up and Q&A

• Summary of key takeaways
• Resources for further learning and software recommendations (FluoroFinder, FlowJoUniversity)
• Open Q&A for specific challenges faced by participants

Before signing up for this nanocourse, please make sure that you can attend all the dates and write them in your calendar. If you do not show up or cancel at very short notice, other interested students will not be able to participate.