Difference between revisions of "Flow cytometry"
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==Principles== | ==Principles== | ||
# Label cells with fluorescent antibodies. | # Label cells with fluorescent antibodies. | ||
# Passed through a channel one-at-a-time (using ''hydrodynamic focusing''<ref>URL: [http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/index.htm http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/index.htm]. Accessed on: 5 August 2010.</ref> | # Passed through a channel one-at-a-time (using ''hydrodynamic focusing''<ref>URL: [http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/index.htm http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/index.htm]. Accessed on: 5 August 2010.</ref>) and interrogated with a laser light. | ||
===Scatter of light=== | |||
Types: | |||
#Forward scatter (FS) -- indicative of cell size. | |||
#Side scatter (SS) -- indicative of cell complexity (granularity). | |||
====Scatter in a table==== | |||
Selected white blood cells & scatter:<ref>URL: [http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/images/blood.gif http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/images/blood.gif]. Accessed on: 5 August 2010.</ref> | |||
{| class="wikitable" | |||
! | |||
! Forward scatter | |||
! Side scatter | |||
|- | |||
| PMNs | |||
| high | |||
| very high | |||
|- | |||
| Monocytes | |||
| high | |||
| moderate/low | |||
|- | |||
| Lymphocytes | |||
| low | |||
| low | |||
|} | |||
==Practical considerations== | |||
*Tissue must be fresh, i.e. cannot be [[formalin]] fixed. | |||
**Viable material may be salvaged from the centre of a large specimen placed in formalin for a short time. | |||
*2 mm x 2 mm x 2 mm piece of tissue is sufficient. | |||
**1 mm<sup>3</sup> is considered the minimum by Mayo Medical Laboratories.<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Specimen/19499 http://www.mayomedicallaboratories.com/test-catalog/Specimen/19499]. Accessed on: 27 December 2012.</ref> | |||
==Interpretation== | |||
:See ''[[Cytometry]]''. | |||
==See also== | ==See also== | ||
Latest revision as of 11:10, 27 December 2012
Flow cytometry is an automated technique used to examine cells that is widely used for the diagnosis and characterization of lymphoma.
Principles
- Label cells with fluorescent antibodies.
- Passed through a channel one-at-a-time (using hydrodynamic focusing[1]) and interrogated with a laser light.
Scatter of light
Types:
- Forward scatter (FS) -- indicative of cell size.
- Side scatter (SS) -- indicative of cell complexity (granularity).
Scatter in a table
Selected white blood cells & scatter:[2]
| Forward scatter | Side scatter | |
|---|---|---|
| PMNs | high | very high |
| Monocytes | high | moderate/low |
| Lymphocytes | low | low |
Practical considerations
- Tissue must be fresh, i.e. cannot be formalin fixed.
- Viable material may be salvaged from the centre of a large specimen placed in formalin for a short time.
- 2 mm x 2 mm x 2 mm piece of tissue is sufficient.
- 1 mm3 is considered the minimum by Mayo Medical Laboratories.[3]
Interpretation
- See Cytometry.
See also
References
- ↑ URL: http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/index.htm. Accessed on: 5 August 2010.
- ↑ URL: http://www.med.umich.edu/flowcytometry/InitialTraining/lessons/lesson1/images/blood.gif. Accessed on: 5 August 2010.
- ↑ URL: http://www.mayomedicallaboratories.com/test-catalog/Specimen/19499. Accessed on: 27 December 2012.
