História da Citometria de Fluxo

O primeiro dispositivo de citometria de fluxo baseado em fluorescência (ICP 11) foi desenvolvido em 1968 por Wolfgang Göhde da Universidade de Münster, Alemanha e foi primeiramente comercializado em 1968/69 pelo empreendedor e fabricante alemão Partec através da Phywe AG em Göttingen. Naquela época, os métodos de absorção ainda eram amplamente favorecidos por outros cientistas em comparação com métodos de fluorescência[1].

O nome original da tecnologia de citometria de fluxo foi citofotometria de pulso (em alemão: mpulszytophotometrie). Apenas 10 anos depois, em 1978, na Conferência da American Engineering Foundation em Pensacola, Flórida, o nome foi alterado para citometria de fluxo, um termo que rapidamente se tornou popular. Logo depois, foram desenvolvidos instrumentos de citometria de fluxo, incluindo o Cytofluorograph (1971) da Bio/Physics Systems Inc. (mais tarde: Ortho Diagnostics), o PAS 8000 (1973) da Partec, o primeiro instrumento FACS da Becton Dickinson (1974), o ICP 22 (1975) da Partec/Phywe e os Epics da Coulter (1977/78) [[2].

Definitions

Flow Cytometry (cyto=cell) (metry=measurement)
Measuring properties of cells in a flowing system

 

Flow Sorting
Sorting (physically separating) cells based on properties measured in a flowing system

 

Flow Cytometer
Early flow cytometers were, in general, experimental devices, but recent technological advances have created a considerable market for the instrumentation, as well as the reagents used in analysis, such as fluorescently-labeled antibodies and analysis software. Modern instruments usually have multiple lasers and fluorescence detectors.

Basic Principle

A beam of light (usually laser light) of a single wavelength is directed onto a hydrodynamically-focused stream of fluid. A number of detectors are aimed at the point where the stream passes through the light beam: one in line with the light beam (Forward Scatter or FSC) and several perpendicular to it (Side Scatter (SSC) and one or more fluorescent detectors).

 

Each suspended particle from 0.2 to 150 micrometers passing through the beam scatters the light in some way, and fluorescent chemicals found in the particle or attached to the particle may be excited into emitting light at a longer wavelength than the light source. This combination of scattered and fluorescent light is picked up by the detectors, and, by analyzing fluctuations in brightness at each detector (one for each fluorescent emission peak), it is then possible to derive various types of information about the physical and chemical structure of each individual particle.

References

  1. "Wallace H. Coulter 1913-1998". Beckman Coulter, Inc. http://www.beckmancoulter.com/hr/ourcompany/oc_WHCoulter_bio.asp. Retrieved on 2008-07-31.
  2. "Joseph R. Coulter Jr. 1924-1995". Beckman Coulter, Inc. http://www.beckmancoulter.com/hr/ourcompany/oc_JRCoulter_bio.asp. Retrieved on 2008-07-31.
  3. Presentation by Bob Auer - 2 December 2008 - Images and text.
  4. Kamentsky, Proceedings of the Conference "Cytology Automation" in Edinburgh, 1970
  5. Images of non Beckman Coulter equipment are from J. Paul Robinson lectures posted on the Purdue website.


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