Cellemme Stephanie L, Van Vorst Matthew, Paramore Elisha, Elliott Gloria D
Department of Mechanical Engineering and Engineering Sciences, University of North Carolina at Charlotte , Charlotte, North Carolina.
Biopreserv Biobank. 2013 Oct;11(5):278-84. doi: 10.1089/bio.2013.0024.
Our prior work has shown that microwave processing can be effective as a method for dehydrating cell-based suspensions in preparation for anhydrous storage, yielding homogenous samples with predictable and reproducible drying times. In the current work an optimized microwave-based drying process was developed that expands upon this previous proof-of-concept. Utilization of a commercial microwave (CEM SAM 255, Matthews, NC) enabled continuous drying at variable low power settings. A new turntable was manufactured from Ultra High Molecular Weight Polyethylene (UHMW-PE; Grainger, Lake Forest, IL) to provide for drying of up to 12 samples at a time. The new process enabled rapid and simultaneous drying of multiple samples in containment devices suitable for long-term storage and aseptic rehydration of the sample. To determine sample repeatability and consistency of drying within the microwave cavity, a concentration series of aqueous trehalose solutions were dried for specific intervals and water content assessed using Karl Fischer Titration at the end of each processing period. Samples were dried on Whatman S-14 conjugate release filters (Whatman, Maidestone, UK), a glass fiber membrane used currently in clinical laboratories. The filters were cut to size for use in a 13 mm Swinnex(®) syringe filter holder (Millipore(™), Billerica, MA). Samples of 40 μL volume could be dehydrated to the equilibrium moisture content by continuous processing at 20% with excellent sample-to-sample repeatability. The microwave-assisted procedure enabled high throughput, repeatable drying of multiple samples, in a manner easily adaptable for drying a wide array of biological samples. Depending on the tolerance for sample heating, the drying time can be altered by changing the power level of the microwave unit.
我们之前的工作表明,微波处理作为一种对基于细胞的悬浮液进行脱水以准备无水储存的方法是有效的,可产生具有可预测和可重复干燥时间的均匀样品。在当前工作中,开发了一种优化的基于微波的干燥工艺,该工艺在先前的概念验证基础上进行了扩展。使用商用微波炉(CEM SAM 255,北卡罗来纳州马修斯)可在可变的低功率设置下进行连续干燥。用超高分子量聚乙烯(UHMW-PE;伊利诺伊州莱克福里斯特的格雷杰公司)制造了一个新的转盘,以便一次干燥多达12个样品。新工艺能够在适合样品长期储存和无菌复水的容器装置中对多个样品进行快速同时干燥。为了确定微波腔内样品干燥的重复性和一致性,对一系列浓度的海藻糖水溶液进行特定时间间隔的干燥,并在每个处理周期结束时使用卡尔费休滴定法评估水分含量。样品在沃特曼S-14共轭释放滤膜(英国梅德斯通的沃特曼公司)上干燥,该滤膜是目前临床实验室使用的玻璃纤维膜。将滤膜裁剪成合适尺寸,用于13毫米Swinnex(®)注射器滤膜支架(马萨诸塞州比勒里卡的密理博(™)公司)。40微升体积的样品通过在20%功率下连续处理可脱水至平衡水分含量,样品间重复性极佳。微波辅助程序能够以易于适应干燥多种生物样品的方式对多个样品进行高通量、可重复的干燥。根据样品加热的耐受性,可通过改变微波装置的功率水平来改变干燥时间。