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  • Title: Characterizing submicron vesicles with wavelength-resolved fluorescence in flow cytometry.
    Author: Fuller RR, Sweedler JV.
    Journal: Cytometry; 1996 Oct 01; 25(2):144-55. PubMed ID: 8891444.
    Abstract:
    Individual synthetic vesicles 0.1-1.0 micron 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 (3 sigma) less than 6.0 x 10(3) and 1.0 x 10(4) 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.
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