History of Flow Cytometry

The first fluorescence-based flow cytometry device (ICP 11) was developed in 1968 by Wolfgang Göhde from the University of Münster, Germany and first commercialized in 1968/69 by German developer and manufacturer Partec through Phywe AG in Göttingen. At that time, absorption methods were still widely favored by other scientists over fluorescence methods1.

The original name of the flow cytometry technology was pulse cytophotometry (German: Impulszytophotometrie). Only 10 years later in 1978, at the Conference of the American Engineering Foundation in Pensacola, Florida, the name was changed to flow cytometry, a term that quickly became popular. Soon after, flow cytometry instruments were developed, including the Cytofluorograph (1971) from Bio/Physics Systems Inc. (later: Ortho Diagnostics), the PAS 8000 (1973) from Partec, the first FACS instrument from Becton Dickinson (1974), the ICP 22 (1975) from Partec/Phywe and the Epics from 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.