Abstract

Individual synthetic vesicles 0.1-1.0 μm in diameter are sized by using single-particle fluorescence emission spectra obtained in a custom sheath flow cell with an imaging spectrograph and a charge-coupled device. Data are acquired at 1 Hz, with limits of detection (3σ) less than 6.0 x 103 and 1.0 x 104 molecules of free sulforhodamine 101 and fluorescein, respectively, and with a spectral range for fluorescence emission collection from 350 to 800 nm (0.45 nm/pixel resolution). The system is used for small-particle population analysis by analyzing a suspension of submicron, unilamellar, synthetic vesicles prepared by standard procedures with phosphatidylserine and Texas red- or fluorescein head-group-conjugated dihydropalmitoylphosphatidylethanolamine. The submicron particles are individually identified, sized, and discriminated based on single-particle fluorescence emission spectra. Excellent agreement is found between fluorescence sizing data and transmission electron microscopic measurements.

Original languageEnglish (US)
Pages (from-to)144-155
Number of pages12
JournalCytometry
Volume25
Issue number2
DOIs
StatePublished - Oct 1 1996

Keywords

  • Charge coupled device
  • Emission spectra
  • Flow cytometry
  • Fluorescence spectroscopy
  • Laser-induced fluorescence
  • Liposomes
  • Particle size distribution
  • Vesicles

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

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