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Sheath fluid control to permit stable flow in rapid mix flow cytometry
L C Seamer1, F Kuckuck, L A Sklar
1University of New Mexico, Cancer Research and Treatment Center, Albuquerque 87131, USA.
Cytometry
|November 30, 1999
Summary
Reducing sheath flow during sample introduction in flow cytometry enables stable laminar flow recovery in under 200 msec. This technique enhances kinetic and high-throughput flow cytometric analysis.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Cell Biology
Background:
- Flow cytometry is valuable for analyzing ligand binding, cell response, and molecular assembly kinetics.
- Historically, flow cytometry was limited to reaction times >5s due to challenges in sample introduction, mixing, and stable flow.
- Automated syringes enable sub-second sample injection, but rapid introduction disrupts laminar flow.
Purpose of the Study:
- To investigate if reducing sheath flow during sample introduction can accelerate stable flow recovery.
- To determine the feasibility of rapid kinetic analysis in flow cytometry.
Main Methods:
- Programmable syringes and valves controlled sample mixing and delivery.
- Stream-in-air detection was used.
- Stable flow recovery was monitored via mean particle fluorescence during sample introduction.
Main Results:
- Without sheath flow reduction, stable flow recovery took over 1 second.
- Reducing sheath flow during the 300 msec sample introduction period allowed stable laminar flow recovery within 200 msec.
Conclusions:
- Automated syringes offer robust sample handling for flow cytometry.
- This method significantly improves sample handling for kinetic and high-throughput analyses.