公开(公告)号：US20070117158A1

公开(公告)日：2007-05-24

申请号：US11434321

申请日：2006-05-12

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

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

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

CPC classification number: C12M41/36 ,  G01N15/1475

Abstract: 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.

Abstract translation: 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。