## Abstract Fluorescence activated cell sorting, FACS, is a widely used method to sort subpopulations of cells to high purities. To achieve relatively high sorting speeds, FACS instruments operate by forcing suspended cells to flow in a single file line through a laser(s) beam(s). Subsequently, thi
Stability of the breakoff point in a high-speed cell sorter
β Scribed by Timothy W. Petersen; Ger van den Engh
- Publisher
- John Wiley and Sons
- Year
- 2003
- Tongue
- English
- Weight
- 109 KB
- Volume
- 56A
- Category
- Article
- ISSN
- 0196-4763
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β¦ Synopsis
Abstract
Background
Highβspeed jetβinβair cytometric sorting requires knowledge of the time it takes a particle to travel from the laser to the point where the jet breaks into droplets. Variations in this breakoff time will result in poorer yields and poorer sort purities.
Methods
This work examined the physical mechanisms that lead to the break up of the jet into droplets and calculated the stability of the droplet breakoff time relative to physical parameters, which govern the behavior of the jet.
Results
We derived the variations in the breakoff time and found that small variations in the drive frequency, temperature, pressure, and drive amplitude can lead to correspondingly large changes in the breakoff time. We found explicitly that the time it takes the jet to break up is not necessarily correlated with the distance to the breakoff point.
Conclusions
Many highβspeed cell sorters use active means to control the breakoff time. A common method to monitor the breakoff time is to visually monitor the breakoff point. This technique in fact may decrease the sorting purity and efficiency by inadvertently correcting for breakoff time variations. We show explicitly the breakoff time's dependence on a number of physical parameters that can be monitored to increase the stability of the breakoff time. Cytometry Part A 56A:63β70, 2003. Β© 2003 WileyβLiss, Inc.
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