MacDonald R C, MacDonald R I, Menco B P, Takeshita K, Subbarao N K, Hu L R
Department of Biochemistry, Northwestern University, Evanston, IL 60208.
Biochim Biophys Acta. 1991 Jan 30;1061(2):297-303. doi: 10.1016/0005-2736(91)90295-j.
The design and performance of a filter holder which enables convenient preparation of volumes of up to a milliliter of large, unilamellar vesicles formed by extrusion (LUVETs) from multilamellar vesicles (MLVs) are described. The filter holder provides for back-and-forth passage of the sample between two syringes, a design that minimizes filter blockage, eliminates the need to change filters during LUVET preparation and reduces preparation time to a few minutes. Replicas of slam-frozen LUVETs in the electron microscope are unilamellar and reasonably homogeneous with an average diameter close to the pore size of the filters used to extrude them. Extrusion per se does not destabilize the vesicles, which trapped a fluorescent dye only when they were disrupted on freeze-thawing and during the first extrusion when most of the MLVs were apparently converted to LUVETs.
本文描述了一种过滤器支架的设计与性能，该支架能够方便地从多层囊泡（MLV）制备出体积达一毫升的由挤压形成的大单层囊泡（LUVET）。该过滤器支架可使样品在两个注射器之间来回通过，这种设计能最大程度减少过滤器堵塞，无需在制备LUVET过程中更换过滤器，并将制备时间缩短至几分钟。在电子显微镜下，骤冷LUVET的复制品为单层且相当均匀，平均直径接近用于挤压它们的过滤器的孔径。挤压本身不会使囊泡不稳定，囊泡仅在冻融时以及首次挤压（此时大多数MLV明显转化为LUVET）过程中破裂时才会捕获荧光染料。
