https://scholars.lib.ntu.edu.tw/handle/123456789/575363
Title: | Separation and detection of rare cells in a microfluidic disk via negative selection | Authors: | Chen, C.-L. Chen, K.-C. Pan, Y.-C. Lee, T.-P. Hsiung, L.-C. Lin, C.-M. Chen, C.-Y. CHING-HUNG LIN BOR-LUEN CHIANG ANDREW WO |
Issue Date: | 2011 | Publisher: | Royal Society of Chemistry | Journal Volume: | 11 | Journal Issue: | 3 | Start page/Pages: | 474-483 | Source: | Lab on a Chip | Abstract: | Cyto-analysis of rare cells often requires separation and detection with each procedure posing substantial challenges. This paper presents a disk-based microfluidic platform for both procedures via an immunomagnetic negative selection process. The microfluidic platform's unique features include a multistage magnetic gradient to trap labeled cells in double trapping areas, drainage of fluid to substantially shorten detection time, and a bin-like regions to capture target cells to facilitate a seamless enumeration process. Proof-of-concept was conducted using MCF7 as target rare cells (stained with anti-cytokeratin-FITC antibodies) spiked into Jurkat Clone E6-1 non-target cells (labeled with anti-CD45-PE and anti-PE BD magnetic beads). Then, mononuclear cells (MNC) from healthy blood donors were mixed with MCF7s, modeling rare cells, and tested in the disk. Results show a non-linear magnetic coupling effect of the multistage magnet substantially increased the trapping efficacy over that of a single magnet, contributing to the depletion rate of Jurkats, which reaches 99.96%. Detection time is extensively shortened by depletion of 95% of non-cell-containing fluid in the collection area. The average yield of detected MCF7 cells is near-constant 60 ± 10% over a wide range of rarity from 10-3 to 10-6 and this yield also holds for the MCF7/MNC complex mixture. Comparison with autoMACS and BD IMagnet separators revealed the average yield from the disk (60%) is superior to that of autoMACS (37.3%) and BD IMagnet (48.3%). The advantages of near-constant yield, roughly 30 min of operation, an acceptable level of cell loss, and potentially low cost system should aid in cyto-analysis of rare cells. ? 2011 The Royal Society of Chemistry. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-78751510615&doi=10.1039%2fc0lc00332h&partnerID=40&md5=977540ad46b651eb3f46b57ee87e62c3 https://scholars.lib.ntu.edu.tw/handle/123456789/575363 |
ISSN: | 1473-0197 | DOI: | 10.1039/c0lc00332h | SDG/Keyword: | article; cell separation; cytology; human; human cell; immunohistochemistry; immunomagnetic separation; leukemia cell line; magnetism; microfluidics; mononuclear cell; priority journal |
Appears in Collections: | 醫學系 |
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