Abstract
A new single-cell microarray chip was designed and developed to separate and analyze single adherent and non-adherent cancer cells. The single-cell microarray chip is made of polystyrene with over 60,000 microchambers of 10 different size patterns (31–40 µm upper diameter, 11–20 µm lower diameter). A drop of suspension of adherent carcinoma (NCI-H1650) and non-adherent leukocyte (CCRF-CEM) cells was placed onto the chip, and single-cell occupancy of NCI-H1650 and CCRF-CEM was determined to be 79% and 84%, respectively. This was achieved by controlling the chip design and surface treatment. Analysis of protein expression in single NCI-H1650 and CCRF-CEM cells was performed on the single-cell microarray chip by multi-antibody staining. Additionally, with this system, we retrieved positive single cells from the microchambers by a micromanipulator. Thus, this system demonstrates the potential for easy and accurate separation and analysis of various types of single cells.
Highlights
Single-cell analysis has implications for our understanding of higher levels of organization of tissues and organisms and, more importantly, may reveal therapeutic approaches to correct flaws in this organization [1]
It is difficult to detect and analyze circulating tumor cells (CTCs) in the peripheral blood of metastatic cancer patients using flow cytometry [4,5]. This is because CTCs in the peripheral blood of metastatic cancer patients are estimated to occur at a frequency of approximately 1 CTC per 106 –107 peripheral blood cells [6,7]
Cell suspension of NCI-H1650 and CCRF-CEM were dropped onto the chip using onto the chip using a pipette, and the cells settled down by gravitational force and adhered to a pipette, and the cells settled down by gravitational force and adhered to the chip surface
Summary
Single-cell analysis has implications for our understanding of higher levels of organization of tissues and organisms and, more importantly, may reveal therapeutic approaches to correct flaws in this organization [1]. In conventional single-cell analysis systems, flow cytometry is one of the most widely used technologies. This technology is a high-throughput system combined with fluorescent labeling and allows for the quantitative determination of various protein levels in a given cell population [2,3]. Flow cytometry is inadequate for evaluating spatial localization of protein expression within single cells. This system is not sufficiently sensitive to detect extremely low ratios (
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