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Thursday, June 28, 2012

FLow Cytometery: Principle, Applications & Advantages


Flow Cytometry is a technique for identifying and sorting cells and their components (eg: DNA) by staining with a fluorescent dye and detecting the fluorescence by using laser beam illumination.
Flow cytometer is the instrument which is used for flow cytometry.

Flow Cytometry Principle
Flow cytometry involves the analysis of the fluorescence and light scatter properties of single particles (e.g. cells, nuclei, chromosomes) during their passage within a narrow, precisely defined liquid stream.

Cell sorting using Flow Cytometer
• Flow cytometry can be used to select and purify a specific subset of cells within a population
• cell sorting based on physical, biochemical and antigenic traits.

Measurable parameters in Flow Cytometry
  • Volume and morphological complexity of cells
  • Cell pigments such as chlorophyll or phycoerythrin
  • DNA (cell cycle analysis, cell kinetics, proliferation etc.)
  • RNA
  • Chromosome analysis and sorting (library construction, chromosome paint)
  • Protein expression and localization
  • Transgenic products in vivo, particularly the Green fluorescent protein or related fluorescent proteins.
  • Cell surface antigens (Cluster of differentiation (CD) markers).
  • Intracellular antigens (various cytokines, secondary mediators etc.)
  • Nuclear antigens Measurable parameters.
  • Enzymatic activity.
  • pH, intracellular ionized calcium, magnesium, membrane potential.
  • Membrane fluidity.
  • Apoptosis (quantification, measurement of DNA degradation, mitochondrial membrane potential, permeability changes, caspase activity)
  • Cell viability.
  • Monitoring electropermeabilization of cells.
  • Oxidative burst.
  • Characterising multidrug resistance (MDR) in cancer cells.
Flow Cytometry Applications:
Flow cytometry has applications in these fields
  • Molecular biology,
  • Pathology,
  • Immunology
  • Plant biology and
  • Marine biology
  • Protein engineering:
Clinical applications in flow cytometry
  • Immunophenotyping of leukemia and lymphoma
  • Platelet Function Analysis
  • Measurement of the Efficacy of Cancer Chemotherapy
  • Cell Function Analysis
  • Applications in Transfusion Medicine
  • Organ Transplantation and Hematopoietic Cell Therapy
  • Applications in Microbiology
Advantages of Flow Cytometry
  • High speed analysis.
  • Measures single cells.
  • Measures large number of cells.
  • Simultaneous analysis of multiple parameters (up to 20)
  • Identifies small sub-populations.
  • Quantification of fluorescence intensities.
  • Sorting of predefined cell populations.
Flow Cytometer Manufacturers
Flow cytometer is manufactured by many companies, some of the are:

Becton Dickinson Inc

Beckman Coulter Inc

Hudson Robotics, Inc.

Agilent Technologies

FACScan™ BD:

FACScan Flow Cytometer - Becton Dickinson

The FACScan™ system is an automated flow cytometer from Becton Dickinson. It analyzes cells as they pass through a focused laser beam one at a time in a moving fluid stream. A BD FACScan flow cytometer can be used for routine research applications, immunophenotyping, and DNA cell-cycle analysis.
The Becton Dickinson FACScan provides the high performance and quality expected from the leader in cell analysis. It is designed and optimized, as a complete system that includes hardware, software, and reagents.

References:
Flow cytometry: principles and applications by By: Dr. Douaa Moh. Sayed
Wikipedia
A Review and Applications of Flow Cytometry, Daniel Heller