Modulating In Vivo T-cell Activation: 15 Color Immunophenotyping, Cytokine Analysis and Cellular Redistribution
Kelly Lundsten
Thursday, June 6, 2013
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Chapters
Introduction
Seminar Outline
LEAF™ (Low Endotoxin Azide Free) Antibodies
Other Interesting In-Vivo Apps
Protocol
Panel Composition
Types of Fluorescent Molecules
Not all Fluors Are Created Equal
Break it Down: Which Fluors for How Many 'Colors'
Panel Composition
Not all Channels Are Created Equal
Spillover Spreading Matrix
Panel Composition
Path of Analysis
Biexponential Scaling and Spreading Error
Panel Composition
Data
CD3 Downregulation
Splenic DC Distribution
Activated DCs in the Lymph Node
Inverse Relationship between CD69 and ICOS Expression after In-Vivo Activation
ICOS/ Voltage Optimization- Splenic T-cells
VISTA affect on ICOS in Lymph Node T-cells
Error Derived from Spillover
CD44 hi (Active) vs CD62L+ (Naïve) Spleen
CD44 hi (Active) vs CD62L+ (Naïve) on LN
Synopsis of Kinetics in Spleen
Synposis of Kinetics in Lymph Node
Unrealistic Compensation? Balance Voltages
Spillover vs. Compensation
Next Steps?
Thanks to all My Colleagues!
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