公开(公告)号：US20070117158A1

公开(公告)日：2007-05-24

申请号：US11434321

申请日：2006-05-12

申请人： Frank Coumans ,  Jan Greve ,  Frank Modica ,  Leon Terstappen ,  Arian Tibbe ,  John Verrant

发明人： Frank Coumans ,  Jan Greve ,  Frank Modica ,  Leon Terstappen ,  Arian Tibbe ,  John Verrant

IPC分类号： G06T7/20 ,  G01N33/567 ,  C12M1/34

CPC分类号： C12M41/36 ,  G01N15/1475

摘要： The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.

摘要翻译： 通过流式细胞术对流体细胞的计数被广泛应用于许多学科，例如评估不同体液中的白细胞亚群或环境样品，食品和体液中的细菌污染。 对于许多应用，仪器的成本，尺寸和复杂性阻止了更广泛的使用，例如资源贫乏国家的艾滋病毒监测中的CD 4分析。 本文公开的新颖的装置，方法和算法在很大程度上克服了这些限制。 简言之，生物样品中的所有细胞都被荧光标记，但只有靶细胞也被磁性标记。 另外，非磁性标记的细胞在修改的载玻片配置中的存活力被成像。 将标记的样品置于室或铜管中，放置在两个楔形磁体之间，以选择性地将磁性标记的细胞移动到铜管的观察表面。 LED照亮细胞，CCD相机捕获由靶细胞发射的荧光的图像。 用新算法进行的图像分析提供了与原始样品的靶细胞浓度相关的表面上的细胞计数。 紧凑型细胞仪系统提供坚固，经济实惠且易于使用的技术，可用于偏远地区。

公开(公告)号：US09134237B2

公开(公告)日：2015-09-15

申请号：US12181399

申请日：2008-07-29

申请人： Mark Carle Connelly ,  Frank Coumans ,  Steve Gross ,  Mike Kelly

发明人： Mark Carle Connelly ,  Frank Coumans ,  Steve Gross ,  Mike Kelly

IPC分类号： G01N33/574 ,  G01N33/53 ,  C12N5/07 ,  G01N33/566 ,  G01N33/553 ,  G01N21/64 ,  G01N33/50 ,  G01N33/569 ,  G01N33/58

CPC分类号： G01N21/6428 ,  G01N21/6458 ,  G01N33/5091 ,  G01N33/5094 ,  G01N33/569 ,  G01N33/57488 ,  G01N33/582 ,  G01N2021/6441

摘要： A sample of blood containing CTCs, or other cells of interest, is stained with fluorescent markers for image analysis and scanned to identify the presence and location within the cartridge of target cells or subcellular elements. A sample containing desired target cells or subcellular elements is then further processed, in part by photobleaching the sample, so that those same targets may be re-analyzed with additional biomarkers conjugated to the same or different fluorochromes using the same imaging criteria that were used for the initial analysis. The present invention has applications with targets such as circulating epithelial, cells, circulating tumor cells, circulating endothelial cells, leukocytes, lymphocyte subsets, cells containing an organelle or receptor of interest, cellular debris, disrupted cells and their debris, or any other formed element that might be captured and imaged. The invention provides a means to further interrogate individual targets of interest, especially when coupled with genetic analysis such as FISH.

公开(公告)号：US20090029378A1

公开(公告)日：2009-01-29

申请号：US12181399

申请日：2008-07-29

申请人： Mark Carle Connelly ,  Frank Coumans ,  Steve Gross ,  Mike Kelly

发明人： Mark Carle Connelly ,  Frank Coumans ,  Steve Gross ,  Mike Kelly

IPC分类号： C12Q1/68 ,  G01N33/53

CPC分类号： G01N21/6428 ,  G01N21/6458 ,  G01N33/5091 ,  G01N33/5094 ,  G01N33/569 ,  G01N33/57488 ,  G01N33/582 ,  G01N2021/6441

摘要： A sample of blood containing CTCs, or other cells of interest, is stained with fluorescent markers for image analysis and scanned to identify the presence and location within the cartridge of target cells or subcellular elements. A sample containing desired target cells or subcellular elements is then further processed, in part by photobleaching the sample, so that those same targets may be re-analyzed with additional biomarkers conjugated to the same or different fluorochromes using the same imaging criteria that were used for the initial analysis. The present invention has applications with targets such as circulating epithelial, cells, circulating tumor cells, circulating endothelial cells, leukocytes, lymphocyte subsets, cells containing an organelle or receptor of interest, cellular debris, disrupted cells and their debris, or any other formed element that might be captured and imaged. The invention provides a means to further interrogate individual targets of interest, especially when coupled with genetic analysis such as FISH.

摘要翻译： 含有CTC或其他感兴趣的细胞的血液样品用用于图像分析的荧光标记染色并扫描以鉴定靶细胞或亚细胞元件的盒内的存在和位置。 然后进一步处理含有所需靶细胞或亚细胞元件的样品，部分通过光漂白样品进行处理，使得可以使用与用于相同或不同荧光染料的相同成像标准的附加生物标志物重新分析相同的靶标 初步分析。 本发明具有诸如循环上皮细胞，细胞，循环肿瘤细胞，循环内皮细胞，白细胞，淋巴细胞亚群，含有感兴趣的细胞器或受体的细胞，细胞碎片，破裂的细胞及其碎片或任何其它形成的元件 这可能被捕获和成像。 本发明提供了进一步询问感兴趣的个体目标的方法，特别是当与FISH等遗传分析相结合时。

公开(公告)号：US20070153280A1

公开(公告)日：2007-07-05

申请号：US11701765

申请日：2007-02-02

申请人： Frank Coumans ,  Arthur Marlin ,  Frank Modica ,  John Verrant

发明人： Frank Coumans ,  Arthur Marlin ,  Frank Modica ,  John Verrant

IPC分类号： G01N21/25

CPC分类号： G01N21/6458 ,  G01N15/1475 ,  G01N2015/1006 ,  G01N2015/1488 ,  G01N2021/6421

摘要： The present invention provides a system for imaging circulating tumor cells from blood after enrichment. The system is designed to provide optimum use in a clinical laboratory setting with minimum laboratory bench top space. Operator intervention is minimized compared to other analytical methodologies. The system is useful in the enumeration and identification of target cells in a sample specimen for screening and detection of early stage pre-metastatic cancer, monitoring for disease remission in response to therapy and selection of more effective dose regimens or alternative therapies for individual patients.

摘要翻译： 本发明提供了一种用于在浓缩后从血液中循环肿瘤细胞成像的系统。 该系统设计为在临床实验室设置中提供最佳使用，具有最少的实验室台面空间。 与其他分析方法相比，操作员干预最小化。 该系统在用于筛选和检测早期转移前癌症的样品标本中的靶细胞的计数和鉴定中是有用的，响应于治疗监测疾病缓解和选择更有效的剂量方案或针对个体患者的替代疗法。