Issue 3, 2011

Separation and detection of rare cells in a microfluidic disk via negative selection

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.

Graphical abstract: Separation and detection of rare cells in a microfluidic disk via negative selection

Supplementary files

Article information

Article type
Paper
Submitted
24 Aug 2010
Accepted
15 Oct 2010
First published
18 Nov 2010

Lab Chip, 2011,11, 474-483

Separation and detection of rare cells in a microfluidic disk via negative selection

C. Chen, K. Chen, Y. Pan, T. Lee, L. Hsiung, C. Lin, C. Chen, C. Lin, B. Chiang and A. M. Wo, Lab Chip, 2011, 11, 474 DOI: 10.1039/C0LC00332H

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