Characterizing submicron vesicles with wavelength-resolved fluorescence in flow cytometry
| Autor: | Robert R. Fuller, Jonathan V. Sweedler |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
education.field_of_study
Materials science Population Biophysics Analytical chemistry Texas Red Cell Biology Hematology Sulforhodamine 101 Fluorescence Fluorescence spectroscopy Pathology and Forensic Medicine chemistry.chemical_compound Endocrinology chemistry Fluorescence cross-correlation spectroscopy Emission spectrum Laser-induced fluorescence education |
| Zdroj: | Cytometry. 25:144-155 |
| ISSN: | 1097-0320 0196-4763 |
| DOI: | 10.1002/(sici)1097-0320(19961001)25:2<144::aid-cyto3>3.0.co;2-h |
| Popis: | 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 × 103 and 1.0 × 104 molecules of free sulforhodamine 101 and fluorescein, respectively, and with a spectal 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 phospatidylserine 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. © 1996 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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