Introduction to Flow Cytometry

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Flow cytometry is a technique that can quantitatively measure (1) protein expression levels per single cell and (2) amounts of different cell-types (based on a protein-maker) for thousands of cells in minutes! Flow cytometers use hydrodynamic focusing to force a mixture of stained cells into a single-file line through a flow cell. Then, each cell, sequentially, passes through a laser beam which excites the fluorophore allowing quantification of protein expression for all proteins that are stained!

In addition, if your cells are alive/surface-stained,* Fluorescence-Activated-Cell Sorting (FACS) instruments are able to use electromagnets to deflect/sort single-cell droplets into different collection tubes. In this manner you can sort a specific cell-type from complex tissue dissection/homogenization. Pretty cool huh!

*NOTE: you can also stain the inside of cells but only by fixing/killing them first

 

REFERENCES:

  1. Howard, G.C.; Kaser, M.R. Making and Using Antibodies: A Practical Handbook CRC Press 2006
  2. Alberts, B. Molecular Biology of the Cell 5th Ed. Garland Science 2008
  3. Murphy, K. Janeway’s Immunobiology 8th Ed. Garland Science 2012

 

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This work by Eugene Douglass and Chad Miller is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

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