title: Microsieve lab-chip device for rapid enumeration and fluorescence in situ hybridization of circulating tumor cells.
abstract: Herein we present a lab-chip device for highly efficient and rapid detection of circulating tumor cells (CTCs) from whole blood samples. The device utilizes a microfabricated silicon microsieve with a densely packed pore array (10(5) pores per device) to rapidly separate tumor cells from whole blood, utilizing the size and deformability differences between the CTCs and normal blood cells. The whole process, including tumor cell capture, antibody staining, removal of unwanted contaminants and immunofluorescence imaging, was performed directly on the microsieve within an integrated microfluidic unit, interconnected to a peristaltic pump for fluid regulation and a fluorescence microscope for cell counting. The latter was equipped with a dedicated digital image processing program which was developed to automatically categorize the captured cells based on the immunofluorescence images. A high recovery rate of >80% was achieved with defined numbers of MCF-7 and HepG2 cancer cells spiked into human whole blood and filtered at a rapid flow rate of 1 mL min(-1). The device was further validated with blood drawn from various cancer patients (8 samples). The whole process, from sample input to result, was completed in 1.5 h. In addition, we have also successfully demonstrated on-microsieve fluorescence in situ hybridization for single cell molecular analysis. This simple method has great potential to supplant existing complex CTC detection schemes for cancer metastasis analysis.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore138669.
authors.forename: Li Shi
authors.initials: LS
authors.lastname: Lim
authors.forename: Min
authors.initials: M
authors.lastname: Hu
authors.forename: Mo Chao
authors.initials: MC
authors.lastname: Huang
authors.forename: Wai Chye
authors.initials: WC
authors.lastname: Cheong
authors.forename: Alfred Tau Liang
authors.initials: AT
authors.lastname: Gan
authors.forename: Xing Lun
authors.initials: XL
authors.lastname: Looi
authors.forename: Sai Mun
authors.initials: SM
authors.lastname: Leong
authors.forename: Evelyn Siew-Chuan
authors.initials: ES
authors.lastname: Koay
authors.forename: Mo-Huang
authors.initials: MH
authors.lastname: Li
mesh_terms: D002452:Cell Count; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D056945:Hep G2 Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D061986:MCF-7 Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D014407:Tumor Cells, Cultured