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Introduction to Flow Cytometry
Flow cytometry is ideally suited for measurement of single cell suspensions, this powerful technique allows for rapid analysis, generating highly quantitative data. In this seminar we will demystify the cytometer through discussion on how they work and discuss best practices in sample preparation/controls.
Your flow cytometry training journey:
o Theory – Intro to Flow Cytometry –in class session
o Protocol/sample preparation/panel design consultations
o Hands-on training (samples required)
o Data analysis (FlowJo)
o Figure preparation for presentations/thesis/manuscript
Intro to Flow Cytometry Session outline
- Instrumentation (what are the key components that make cytometers work)
- Pitfalls of sample preparation – how do I maximize my signal to noise
- Do I need controls? (Yes, several! We will discuss the different types)
- Resources to further your learning
- How do I select the instrument for my hands-on training?
o Virtual orientation of the instruments in the flow core
o What do I need to prepare for the hands-on training?
- When will I get access to the Flow Core? (RFBMS)
- Rules of the Flow Core
- Short comprehension Quiz
Reference Materials
Andrea Cossarizza , Hyun-Dong Chang , Andreas Radbruch , Sergio Abrignani et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) Eur J Immunol 2021 Dec;51(12):2708-3145
Andrew Reichard, Kewal Asosingh. Best Practices for Preparing a Single Cell Suspension from Solid Tissues for Flow Cytometry. Cytometry A 2019 Feb;95(2):219-226.
Brooks HL, de Castro Brás LE, Brunt KR, et al. Guidelines on antibody use in physiology research. Am J Physiol Renal Physiol. 2024 Mar 1;326(3):F511-F533.
Ruud Hulspas. Titration of Fluorochrome-Conjugated Antibodies for Labeling Cell Surface Markers on Live Cells. Current Protocols in Cytometry 6.29.1-6.29.9, October 2010
Burn OK, Mair F, Ferrer-Font L. Combinatorial antibody titrations for high-parameter flow cytometry. Cytometry A. 2024 May;105(5):388-393.
Zofia Maciorowski, Pratip K Chattopadhyay, Paresh Jain. Basic Multicolor Flow Cytometry Curr Protoc Immunol 2017 Apr 3:117:5.4.1-5.4.38.
Julie Demaret, Morgane Gossez, Fabienne Venet, Guillaume Monneret. Intracellular Flow Cytometry Improvements in Clinical Studies. Methods Mol Biol 2017:1524:315-327.