A microscale sorting cytometer for cell-based screens

用于细胞筛选的微型分选细胞仪

基本信息

批准号：
6759959
负责人：
Joel Voldman
金额：
$ 20.32万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2009-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): We propose the development of a cytometer that combines microscopic imaging with automated sorting. The cytometer addresses two steps in cell-based genetic screens: observation of phenotypes and retrieval of "mutants". Our goal is to be able to sort cells where the sort parameter is based upon the rich information available from microscopy: morphology, intracellular fluorescence, dynamics. Our cytometer is essentially an "active" coverslip that uses an array of reversible addressable traps created with semiconductor technology to hold individual cells in place for observation and release selected cells. The traps use an electrical analogue of optical tweezers--termed dielectrophoresis--to transiently hold the adherent cells in place until they attach to the substrate. After attachment, the cytometer is a passive chip that fits under a conventional upright fluorescence microscope. After live-cell assays have been performed, individual cells in selected traps can be released. The specific aims of our research are to (1) create trapping arrays of increasing complexity, starting with a 4x4 array to verify function, a 20x20 array to characterize sorting, and ending in a 100x100 array for use in an library screen; (2) create the control systems to provide constant temperature, pH, and oxygen tension on-chip coupled with automated microscopy; (3) investigate effects of our cytometer on cell health and perform a phenotype screen.

描述（由申请人提供）：我们提出了将微观成像与自动排序结合的细胞仪的开发。细胞仪介绍了基于细胞的遗传筛选的两个步骤：观察表型和“突变体”的检索。我们的目标是能够根据显微镜可用的丰富信息进行排序参数的细胞：形态，细胞内荧光，动力学。我们的细胞仪本质上是一种“活跃”的盖玻片，它使用了用半导体技术创建的一系列可逆的可寻址陷阱，以将单个单元固定在适当的观察和释放选定的细胞。陷阱使用光学镊子的电类似物 - 近距离介电的电泳 - 瞬时将粘附的细胞固定在适当的位置，直到它们附着在底物上。附着后，细胞仪是一个被动芯片，适合在常规的直立荧光显微镜下。进行活细胞测定后，可以释放选定陷阱中的单个单元格。我们研究的具体目的是（1）创建增加复杂性的捕获阵列，从4x4阵列开始以验证函数，一个20x20阵列，以表征分类，并以100x100阵列结束，以在库屏幕中使用；（2）创建控制系统，以在片上提供恒定的温度，pH和氧张力以及自动显微镜；（3）研究我们的细胞仪对细胞健康的影响并执行表型筛选。