Simultaneous Multipass Resistive-Pulse Sensing and Fluorescence Imaging of Liposomes

被引:6
作者
Schmeltzer, Alexandra J. [1 ]
Peterson, Eric M. [2 ]
Harris, Joel M. [1 ]
Lathrop, Daniel K. [2 ]
German, Sean R. [2 ]
White, Henry S. [1 ]
机构
[1] Univ Utah, Dept Chem, Salt Lake City, UT 84112 USA
[2] Elect Biosci Inc, Salt Lake City, UT 84108 USA
基金
美国国家卫生研究院;
关键词
resistive-pulse sensing; Coulter counter; nanopipet; fluorescence; liposomes; cryoTEM; INDIVIDUAL NANOPARTICLES; EXTRACELLULAR VESICLES; SUBMICRON PARTICLES; SURFACE-CHARGE; PORES; FLOW; TRANSPORT; DYNAMICS; SIZE;
D O I
10.1021/acsnano.3c12627
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Simultaneous multipass resistive-pulse sensing and fluorescence imaging have been used to correlate the size and fluorescence intensity of individual E. coli lipid liposomes composed of E. coli polar lipid extracts labeled with membrane-bound 3,3-dioctadecyloxacarbocyanine (DiO) fluorescent molecules. Here, a nanopipet serves as a waveguide to direct excitation light to the resistive-pulse sensing zone at the end of the nanopipet tip. Individual DiO-labeled liposomes (>50 nm radius) were multipassed back and forth through the orifices of glass nanopipets' 110-150 nm radius via potential switching to obtain subnanometer sizing precision, while recording the fluorescence intensity of the membrane-bound DiO molecules. Fluorescence was measured as a function of liposome radius and found to be approximately proportional to the total membrane surface area. The observed relationship between liposome size and fluorescence intensity suggests that multivesicle liposomes emit greater fluorescence compared to unilamellar liposomes, consistent with all lipid membranes of the multivesicle liposomes containing DiO. Fluorescent and nonfluorescent liposomes are readily distinguished from each other in the same solution using simultaneous multipass resistive-pulse sensing and fluorescence imaging. A fluorescence "dead zone" of similar to 1 mu m thickness just outside of the nanopipet orifice was observed during resistive-pulse sensing, resulting in "on/off" fluorescent behavior during liposome multipassing.
引用
收藏
页码:7241 / 7252
页数:12
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