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Related Experiment Videos

Small-volume rapid-mix device for subsecond kinetic analysis in flow cytometry.

Yang Wu1, Gordon Zwartz, Gabriel P Lopez

  • 1Department of Chemical and Nuclear Engineering, University of New Mexico Health Sciences Center, Albuquerque, New Mexico 87131, USA.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|August 6, 2005
PubMed
Summary

This study introduces a new rapid-mix flow cytometry device that precisely delivers and mixes small sample volumes. It enables subsecond kinetic analysis of biological reactions using minimal precious reagents.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Biophysics

Background:

  • Rapid-mix flow cytometry is vital for analyzing ligand binding and cellular responses.
  • Previous advancements focused on sample delivery, neglecting reagent volume needs.
  • Precious biological reagents necessitate low-volume handling methods.

Purpose of the Study:

  • To develop a modular, precisely regulated rapid-mix device for small-volume sample delivery to flow cytometers.
  • To address the volumetric requirements of precious biological reagents in flow cytometry experiments.
  • To enable mechanistic analysis of fast biological reactions with minimal sample input.

Main Methods:

  • A modular rapid-mix device was engineered using programmable syringes, valves, and fluidic components.

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  • The device was tested with a bead-based assay to characterize binding kinetics.
  • Native biotin and fluorescein biotin-bearing beads were used to validate performance.

    • Main Results:

    • The device efficiently mixed and delivered 35- to 45-microliter sample and reagent aliquots.
    • Kinetic data from fluorescein biotin beads were analyzed to calibrate device performance.
    • Effective sample delivery and mixing efficiency were demonstrated.

    Conclusions:

    • The developed rapid-mix device accurately detects subsecond biological reaction kinetics with microliter sample volumes.
    • Device dimensions were minimized, and quantitative sample delivery/analysis were optimized.
    • The modular design allows for adaptation to diverse experimental protocols.