Molter Timothy W, McQuaide Sarah C, Suchorolski Martin T, Strovas Tim J, Burgess Lloyd W, Meldrum Deirdre R, Lidstrom Mary E
University of Washington, Benjamin Hall Research Building, 616 NE Northlake Pl. Rm. 440, Seattle, WA 98105.
Sens Actuators B Chem. 2009 Jan 15;135(2):678-686. doi: 10.1016/j.snb.2008.10.036.
Due to interest in cell population heterogeneity, the development of new technology and methodologies for studying single cells has dramatically increased in recent years. The ideal single cell measurement system would be high throughput for statistical relevance, would measure the most important cellular parameters, and minimize disruption of normal cell function. We have developed a microwell array device capable of measuring single cell oxygen consumption rates (OCR). This OCR device is able to diffusionally isolate single cells and enables the quantitative measurement of oxygen consumed by a single cell with fmol/min resolution in a non-invasive and relatively high throughput manner. A glass microwell array format containing fixed luminescent sensors allows for future incorporation of additional cellular parameter sensing capabilities. To demonstrate the utility of the OCR device, we determined the oxygen consumption rates of a small group of single cells (12 to 18) for three different cells lines: murine macrophage cell line RAW264.7, human epithelial lung cancer cell line A549, and human Barrett's esophagus cell line CP-D.
由于对细胞群体异质性的关注,近年来用于研究单细胞的新技术和方法有了显著增加。理想的单细胞测量系统应具有高通量以确保统计相关性,能测量最重要的细胞参数,并将对正常细胞功能的干扰降至最低。我们开发了一种能够测量单细胞耗氧率(OCR)的微孔阵列装置。这种OCR装置能够通过扩散隔离单个细胞,并以非侵入性且相对高通量的方式,实现对单个细胞以fmol/分钟分辨率的耗氧量进行定量测量。包含固定发光传感器的玻璃微孔阵列形式,为未来纳入额外的细胞参数传感能力提供了可能。为了证明OCR装置的实用性,我们测定了一小群(12至18个)单细胞针对三种不同细胞系的耗氧率:小鼠巨噬细胞系RAW264.7、人上皮肺癌细胞系A549和人巴雷特食管细胞系CP-D。