Contents

• Introduction
• Principles of Flow Cytometry
1. The Flow Cytometer
1. FACS Fluidics system
2. FACS Optics I : Detection – Lasers and Types of Optical Filters
3. FACS Optics II : Processing – and Quantification of Signals
4. Electrostatic Sorting of Cells
2. The Fluorophores
1. The Basis of Fluorescence – Excitation and Emission
2. Common Fluorophores and Spectral Characteristics
3. Selecting the Right Fluorophore Conjugate (Key Properties)
4. Single and Tandem Dyes
5. Spectral Overlap and Compensation
• FACS Data Acquisition and Analysis
1. FACS Data Display – What does FACS data look like?
2. FACS Data Analysis – Gating Strategies
• Sample Preparation and Protocols
1. Preparation of Cells for FACS Staining
1. Preparation of Suspension Culture Cells
2. Preparation of Adherent Culture Cells
3. Preparation of Cryopreserved Cells
4. Preparation of Blood Mononuclear Cells
5. Preparation of Cells from Tissues
2. Common FACS Staining Protocols
1. Direct Immunostaining of Surface Antigens
2. Indirect Immunostaining of Surface Antigens
3. General Immunostaining of Intracellular Antigens – Permeabilization Methods
4. Intracellular Cytokine Stimulation and Phospho-immunostaining
5. Dye Efflux Staining
6. DNA Content and Cell Cycle Analysis
• Optimization Tips
1. Sample Preparation and Cell Quality
2. General Immuno-staining (Fluorescence Staining)
3. Intracellular Staining: Fixation and Permeabilization
4. Multicolor Panel Design
5. Experimental (Staining) Controls
• Troubleshooting Guide
1. Weak Fluorescence Intensity or No Fluorescent Signal
2. Excess Fluorescent Signal
3. High Background or Non-specific Staining
4. High Background Scatter or Abnormal Scatter Profile of Cells
5. Abnormal Event Rate
6. Loss of Epitope
• FAQs
• Ordering Information