384-Channel parallel microfluidic cytometer for rare-cell screening
384-Channel parallel microfluidic cytometer for rare-cell screening
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DOI：
10.1039/b811889b
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发表时间：
2009-01-01
期刊：
影响因子：
6.1
通讯作者：
Ehrlich, Daniel J.
中科院分区：
文献类型：
--
作者：
Mckenna, Brian K.;Selim, A. A.;Ehrlich, Daniel J.
我们构建了一台384通道并行微流控细胞仪(PMC)。多通道体系结构允许在大约6-10分钟内读出基于细胞的屏幕的384个独特样本，大约是传统荧光激活细胞仪系统(FACS)的30倍。该体系结构还允许信号积分时间在比单通道FAC的实际范围更大的范围内变化，并且适合于在高负片背景中检测稀有细胞。通过使用该系统对羧基末端甲状旁腺激素受体(CPTHR)表达克隆筛查中最困难的早期阶段的罕见克隆性骨细胞进行计数，证实了信噪比优势。这个问题需要在一百万张底片的背景中找到几十个阳性细胞。该系统是围绕扫描激光共聚焦探测器和96尖端机器人移液器实现自动化的，可以在384孔板中保持体外培养系统。因此，利用现有的高通量培养设施直接用于生物学应用是可行的。PMC系统适合于高样本数量的细胞学，具有非凡的时间同步和稀有细胞灵敏度能力。有限的处理大样本数量的能力限制了单通道流式细胞仪在组合细胞分析中的应用，因此PMC在高通量筛选中可能有重要的应用。
We have constructed a 384-channel parallel microfluidic cytometer (PMC). The multichannel architecture allows 384 unique samples for a cell-based screen to be read out in approximately 6-10 min, about 30-times the speed of a conventional fluorescence-activated cytometer system (FACS). This architecture also allows the signal integration time to be varied over a larger range than is practical in single-channel FACS and is suitable for detection of rare-cells in a high background of negatives. The signal-to-noise advantages have been confirmed by using the system to count rare clonal osteocytes in the most difficult early stages of an expression-cloning screen for the carboxy- terminal parathyroid hormone receptor (CPTHR). This problem requires finding several dozen positive cells in a background of one million negatives. The system is automated around a scanning laser confocal detector and a 96-tip robotic pipettor and can maintain in vitro cultures on-system in 384-well plates. It is therefore directly practical for biology applications using existing high-throughput culture facilities. The PMC system lends itself to high-sample-number cytometry with an unusual capability for time synchronization and rare-cell sensitivity. A limited ability to handle large sample numbers has restricted applications of single-channel FACS in combinatorial cell assays; therefore the PMC could have a significant application in high-throughput screening.