Transcription of Flow Cytometry Basics Guide - Bio-Rad
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Flow Cytometry Basics Guide - Bio-Rad
Flow CytometryPractical Advice to Get You Started in FlowFlow Cytometry Basics GuideVisit for more information Flow Cytometry Basics Guide | 1 Table of ContentsChapter 1 Principles of the Flow CytometerFluidics System ..3 Optics and Detection ..4 Signal and Pulse Processing ..6 Electrostatic Cell Sorting ..9 Chapter 2 Principles of FluorescenceFluorophores and Light ..11 Fluorescence ..12 Why Use a Fluorescent Marker? ..13 Which Fluorophores are Useful for Flow Cytometry ? ..13 single and Tandem Dyes ..14 Fluorescent Proteins ..14 Fluorescence Compensation ..16 Compensation Controls ..18 Chapter 3 Data AnalysisGates and Regions ..19 single -Parameter or Univariate Histograms ..21 Two-Parameter or Bivariate Histograms ..22 Backgating to Confirm Gating Strategies ..24 Chapter 4 Controls in Flow cytometryUnstained Controls ..25 Isotype Controls ..26 single Staining and Compensation Controls ..27 Fc Blocking Controls.
Fig. 1. Hydrodynamic focusing produces a single stream of particles. Without hydrodynamic focusing, the cuvette (typically 250 x 250 µm or 180 x 480 µm) or nozzle of the instrument (typically 70-130 µm) would not create a focused stream of cells and analysis of single cells would not be possible . With hydrodynamic focusing the cells flow in
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