DESIGNABLE REGULATION OF CELL FILTRATION THROUGH MICROPORE ARRAY
Yaoping Liu1,2, Xiaolong Rao3, Wengang Wu1,4,5, Yan Sun2,6, Wei Wang1,2,5
1Institute of Microelectronics, Peking University, China;
2State Key Laboratory of Transducer Technology, Chinese Academy of Sciences, China;
3Peking University First Hospital, China;
4National Key Laboratory of Science and Technology on Micro/Nano Fabrication, China;
5Innovation Center for Micro-Nano-electronics and Integrated System, China;
6Beihang University, China
Micropore-arrayed filter has been considered as the only technique for rare cell separation from a large volume of clinical sample (>10 mL) with a high recovery rate. However, all the present filtration was designed based on simply comparing the geometric sizes of the target cells and the micropore. As cells usually have a broad diameter distribution, the purity of the target cell after filtration is usually questionable, especially when the diameter distributions of target and background cells are overlapped. This work reports a designable regulation of cell filtration through the microporearrayed filter by tuning the cell deformability with cytochalasin D.
Cytochalasin D was thought to have defined effects on cytoskeleton depolymerization and actin aggregation. Cells treated with cytochalasin D had an increased deformability and passed through the micropore even if which is of smaller size. The effects of concentrations (0.5, 1.0, 2.0, 5.0, 10 μg/ml for 30 min) and durations (5, 10, 20, 30, 40 min at 5.0 μg/ml) on Hela cells were studied. Hela cells from solution before filtration and filtrate after filtration were imaged under a microscopy and diameters were extracted via Image J.
The decreased quantity and intensity of green fluorescence from filament with the increments of concentration and duration proved that cytochalasin D promoted the cytoskeleton depolymerization, which contributed to the deformability increase and caused the diameter distributions of cells after filtration to approach to those before filtration. Besides, the viability of cytochalasin D-treated cells was tested with trypan blue staining and were higher than 94%. This paper demonstrated a designable regulation of cell filtration through micropore-arrayed filter, which will be useful to establish an actively modulating method to facilitate a highly efficient rare cell separation with a high purity