Introduction to Flow Cytometry
In this seminar we will demystify the cytometer through discussion on how they work and discuss best practices in sample preparation/controls.
Intro to Flow Cytometry Session-Part I
-Instrumentation (what are the key components that make cytometers work)
-Pitfalls of sample preparation – how do I maximize my signal to noise
-Panel design
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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
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