Introduction to FlowCytometry

What Is Flow Cytometry ?

 Flow= the motion characteristics of fluids.

Cyto=cells

metry=measurement

 

Cytometry

is a general name for a group of biological methods used to measure various parameters of cells. Parameters which can be measured by cytometric methods are cell size, the stage of the cell cycle, the DNA content of the cell, the existence or absence of specific proteins on the cell surface or in the cytoplasm, to name but a few. (Wikipedia)

 

Some applications of flow cytometry

• DNA/Cell Cycle analysis 

• Cell viability

• Cell proliferation

• Intracellular ionic (e.g. Ca2+ ) fluxes

• Multicolor phenotyping (cell surface)

• Multicolor phenotyping (intracellular)

• Monocyte oxidative burst

• Monocyte phagocytosis

• Neutrophil oxidative burst

• Neutrophil phagocytosis

• Microbiological analysis

• Cell trafficking

• Cellular and antibody or complementmediated cytotoxicity

• Sorting on the basis of morphology (FSC or SSc) and/or fluorescent characteristics

 

What does a Flow Cytometer do?

Analyses light signals to determine: Phenotype and Function 

 

What’s inside a Flow Cytometer ?

 3 core systems – fluidics, optics, electronics

FlowCytometry systems

 

 What type of signals do we see with Flow ?

• Scatter – Forward Scatter (FSC)

• parallel or Perpendicular FSC – Side scatter (SSC)

• Fluorescence – FITC , PE, APC, GFP, DAPI (plus lots lots more)