Fuller R R, Sweedler J V
Department of Chemistry and Beckman Institute, University of Illinois, Urbana, USA.
Cytometry. 1996 Oct 1;25(2):144-55. doi: 10.1002/(SICI)1097-0320(19961001)25:2<144::AID-CYTO3>3.0.CO;2-H.
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.
通过使用在配备成像光谱仪和电荷耦合器件的定制鞘流池内获得的单颗粒荧光发射光谱，对直径为0.1 - 1.0微米的单个合成囊泡进行尺寸测定。数据采集频率为1 Hz，游离磺基罗丹明101和荧光素的检测限（3σ）分别小于6.0×10³和1.0×10⁴分子，荧光发射采集的光谱范围为350至800 nm（分辨率为0.45 nm/像素）。该系统用于通过分析由标准程序制备的、含有磷脂酰丝氨酸和Texas红或荧光素头部基团共轭二氢棕榈酰磷脂酰乙醇胺的亚微米单层合成囊泡悬浮液，来进行小颗粒群体分析。基于单颗粒荧光发射光谱对亚微米颗粒进行单独识别、尺寸测定和区分。荧光尺寸测定数据与透射电子显微镜测量结果之间具有极佳的一致性。
