Silicon membrane filter designed by fluid dynamics simulation and near-field stress analysis for selective cell enrichment.

Selective cell enrichment technologies can play an important role in both diagnostic and therapeutic areas. However, currently used cell sorting techniques have difficulties in rapidly isolating only the desired target cells from a large volume of body fluids. In this work, we developed a filtering system that can quickly separate and highly concentrate cells from a large volume of solution, depending on their size, using a silicon membrane filter. To overcome the problems caused by material limitations of the brittle silicon, we designed a novel membrane filter with various pore designs. From these designs, the most optimal design with high pore density, while preventing crack formation was derived by applying fluid dynamics simulation and near-field stress analysis. The membrane filter system using the selected design was fabricated, and cell filtration performance was evaluated. The LNCaP cell in horse blood was recovered up to 86% and enriched to 187-fold compared to initial cell populations after filtration at a flow rate of 5 mL/min. The results demonstrate that the filter presented in this study can rapidly and selectively isolate target cells from a large volume of body fluid sample.

SSA-MOA: a novel CTC isolation platform using selective size amplification (SSA) and a multi-obstacle architecture (MOA) filter.

Circulating tumor cells (CTCs) have gained increasing attention as physicians and scientists learn more about the role these extraordinarily rare cells play in metastatic cancer. In developing CTC technology, the critical criteria are high recovery rates and high purity. Current isolation methods suffer from an inherent trade-off between these two goals. Moreover, ensuring minimal cell stress and robust reproducibility is also important for the clinical application of CTCs. In this paper, we introduce a novel CTC isolation technology using selective size amplification (SSA) for target cells and a multi-obstacle architecture (MOA) filter to overcome this trade-off, improving both recovery rate and purity. We also demonstrate SSA-MOA's advantages in minimizing cell deformation during filter transit, resulting in more stable and robust CTC isolation. In this technique, polymer microbeads conjugated with anti-epithelial cell adhesion molecules (anti-EpCAM) were used to selectively size-amplify MCF-7 breast cancer cells, definitively differentiating from the white blood cells (WBCs) by avoiding the size overlap that compromises other size selection methods. 3 µm was determined to be the optimal microbead diameter, not only for size discrimination but also in maximizing CTC surface coverage. A multi-obstacle architecture filter was fabricated using silicon-on-glass (SOG) technology-a first such application of this fabrication technique-to create a precise microfilter structure with a high aspect ratio. The filter was designed to minimize cell deformation as simulation results predicted that cells captured via this MOA filter would experience 22% less moving force than with a single-obstacle architecture. This was verified by experiments, as we observed reliable cell capture and reduced cell deformation, with a 92% average recovery rate and 351 peripheral blood leukocytes (PBL) per millilitre (average). We expect the SSA-MOA platform to optimize CTC recovery rates, purity, and stability, increasing the sensitivity and reliability of such tests, thereby potentially expanding the utilization of CTC technologies in the clinic.