Flow cytometry basics for the non-expert /
Saved in:
| Author / Creator: | Goetz, Christine, author. |
|---|---|
| Imprint: | Cham : Springer, 2018. |
| Description: | 1 online resource (229 pages) |
| Language: | English |
| Series: | Techniques in Life Science and Biomedicine for the Non-Expert Techniques in life science and biomedicine for the non-expert. |
| Subject: | |
| Format: | E-Resource Book |
| URL for this record: | http://pi.lib.uchicago.edu/1001/cat/bib/11737611 |
Table of Contents:
- Intro; Preface; Acknowledgments; Contents; About the Book; Chapter 1: Flow Cytometry: Definition, History, and Uses in Biological Research; 1 Components of Flow Cytometry; 2 Definition of Flow Cytometry; 3 History of Flow Cytometry; 3.1 Cellular Impedance and the Coulter Principle; 3.2 Fluorescence-Based Flow Cytometers; 3.3 Monoclonal and Polyclonal Antibodies; 3.4 Fluorescent Molecules; 3.5 Fluorescence Activated Cell Sorting (FACS); 4 Modern Flow Cytometers; 4.1 Fluidic-Based Flow Cytometers; 4.2 Acoustic-Based Flow Cytometers; 4.3 Mass Cytometers (CyTOF); References.
- Chapter 2: Physics of a Flow Cytometer1 Components of Fluidic-Based Flow Cytometers; 1.1 Fluidics; 1.2 Optics; 1.3 Electronics; 2 Putting It All Together; 3 Materials; 3.1 Flow Antibodies; 3.2 Cell Activation Reagents; 3.3 Cell Purification Kits; 3.4 Staining Buffers/Fc Block Reagents; References; Chapter 3: The Language of Flow Cytometry and Experimental Setup; 1 Flow Data Plots and Gating; 1.1 Flow Data Plots; 1.2 Gating; 2 Mean Fluorescence Intensity (MFI) and Biexponential vs. Logarithmic Scaling; 2.1 Mean Fluorescence Intensity (MFI); 2.2 Logarithmic vs. Biexponential Scaling.
- 3 Forward Scatter (FSC)/Side Scatter (SSC) and Doublet Exclusion3.1 FSC/SSC; 3.2 Doublet Exclusion; 4 CS & T and Basic Experimental Setup; 4.1 CS 4.2 Basic Experimental Setup; 4.2.1 Experimental/Flow Panel Design; 4.2.2 Surface and Intracellular Staining on iTregs; 4.2.3 Experimental Setup on the Fortessa; 4.2.4 Sample Acquisition and Analysis; 5 Basic Concepts of Compensation; 6 Materials; 6.1 Flow Antibodies; 6.2 Cell Activation Reagents; 6.3 Cell Purification Kits; 6.4 Staining Buffers/Fc Block Reagents; References; Chapter 4: Fluorochrome Choices for Flow Cytometry.
- 1 Fluorochromes and Their Pros and Cons2 Spectrum Viewers and How to Use Them; 3 Fluorochrome Combinations to Use/Avoid Together; 4 Laser Strength on the Fortessa vs. the Calibur; 5 High- vs. Low-Density Cell Markers and Fluorochrome Choice; 6 Materials; 6.1 Flow Antibodies; 6.2 Staining Buffers/Fc Block Reagents; References; Chapter 5: Using the Process of Compensation to Prevent False Positive Data Caused by Fluorescence Spillover: A Practical Example; 1 Compensation: A Practical Example; 2 Manual Compensation by Comparing the Median Fluorescence Intensity of Cell Populations.
- 3 Considerations When Performing Compensation3.1 Using the Same Compensation Settings for Multiple Experiments; 3.2 Dimming of Fluorochromes as a Result of Compensation; 3.3 Consider the Number of Overlapping Fluorochromes; 3.4 Consider If Compensation Is Necessary; 3.5 Consider If Fluorochromes with High Spectral Overlap Can Be Separated on Two Different Cell Types; 3.6 Using Cells as Single-Stain Controls vs. Beads; 4 Materials; 4.1 Flow Antibodies; 4.2 Staining Buffers/Fc Block Reagents; References; Chapter 6: Primary and Secondary Antibodies and Flow Cytometry Controls.