Liposome Sterile Filtration Characterization via X-ray Computed Tomography and Confocal Microscopy.
Liposome Sterile Filtration Characterization via X-ray Computed Tomography and Confocal Microscopy.
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DOI：
10.3390/membranes11110905
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发表时间：
2021-11-22
期刊：
影响因子：
4.2
通讯作者：
Bracewell DG
中科院分区：
文献类型：
--
作者：
Johnson TF;Jones K;Iacoviello F;Turner S;Jackson NB;Zourna K;Welsh JH;Shearing PR;Hoare M;Bracewell DG
应用两种高分辨率的三维成像技术对脂质体的灭菌级双层过滤进行可视化和表征，使膜结构与功能和性能相关。用两种标称截留率分别为650 nm和200 nm的聚醚砜膜过滤平均直径为143 nm、多分散指数为0.1的脂质体。对包括差压在内的操作条件进行了评估。像素尺寸为63 nm的X射线计算机断层扫描能够分辨每个膜的内部几何形状。可以测量上游和下游膜的各自的不对称性和对称性，使用孔网络模型来识别作为通过成像体积的距离的函数的孔大小。重建的三维数字数据集是模拟每种多孔结构的曲折流动的基础。共聚焦显微镜显示使用荧光染料的脂质体在每一膜内的滞留，还进行了细菌挑战。结果表明，当压降从0.07 Mpa增加到0.21 Mpa时，上游膜的荧光截留率不同。这些结果突出了补充成像方法加深对脂质体灭菌分级过滤的理解的能力。
Two high resolution, 3D imaging techniques were applied to visualize and characterize sterilizing grade dual-layer filtration of liposomes, enabling membrane structure to be related with function and performance. Two polyethersulfone membranes with nominal retention ratings of 650 nm and 200 nm were used to filter liposomes of an average diameter of 143 nm and a polydispersity index of 0.1. Operating conditions including differential pressure were evaluated. X-ray computed tomography at a pixel size of 63 nm was capable of resolving the internal geometry of each membrane. The respective asymmetry and symmetry of the upstream and downstream membranes could be measured, with pore network modeling used to identify pore sizes as a function of distance through the imaged volume. Reconstructed 3D digital datasets were the basis of tortuous flow simulation through each porous structure. Confocal microscopy visualized liposome retention within each membrane using fluorescent dyes, with bacterial challenges also performed. It was found that increasing pressure drop from 0.07 MPa to 0.21 MPa resulted in differing fluorescent retention profiles in the upstream membrane. These results highlighted the capability for complementary imaging approaches to deepen understanding of liposome sterilizing grade filtration.
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