Method
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Notes
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Applicability
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Adaptability |
Flow path:
- Open space
- flow cell (quartz, plastic)
- Micro fluidics, microchip. Fishman-R |
Most bench top flowcytometers use open space path. Flowcytometers for toxic and hazardous substances, and oceanographic use typically enclosed path.
|
Design of flow path may affect optical performance, sensitivity
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Maintenance:
- flow path needs to be easily accessible for cleaning or replacement
Catastrophic failure (flooding)
- flow path needs to be enclosed
|
Physical volume vs. detectable volume ("virtual core") - SeaFlow, Sealable
|
Ability to re-process primary data. |
|
Pros:
- Allows processing of undiluted sample
- no need for sheath fluid
Cons:
- virtual core is harder to characterize then physical volume
|
| Sheath fluid, vs. undiluted sample |
|
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|
fluorescent and calibration bits.
|
Used for calibration on some (most) benchtop flowcytometers |
|
Cons:
- hard to implement in "in situ" instrument
|
Cell sorting:
- Mechanical sorting into "catcher tube". Enclosed path, one population.
- Electrostatic deflection |
actively separate and isolate particles having specified properties. |
seem to be complex to implement and currently does not required
|
|
Focusing:
- Hydrodynamic focusing: coaxial laminar flow
- Acoustic focusing (Ward) |
This is physical sample focusing
|
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| Real time or post processing, or both |
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Real time processing: required
Post processing: beneficial
|
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Labeling
- Fluorescent
- Staining (Worden - bacterial communities)
- Isotope labeling |
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Processing and software
- Gating and analyzing software
- Population identification using computational methods (rare and hidden populations)
- Population identification (Worden): trigger on scattering signal is used, and then accumulate data with other parameters. This is "see everything" approach |
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