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    Multipass cavity for illumination and excitation of moving objects
    2.
    发明申请
    Multipass cavity for illumination and excitation of moving objects 有权
    多光腔用于照明和激发移动物体

    公开(公告)号：US20040080748A1

    公开(公告)日：2004-04-29

    申请号：US10691292

    申请日：2003-10-21

    Applicant: Amnis Corporation

    Inventor： William E. Ortyn ,  David A. Basiji ,  Keith L. Frost ,  N. Robert McRuer

    IPC: G01N021/00

    CPC classification number: G02B27/126 ,  G01J3/2803 ,  G01J3/2823 ,  G01N15/1434 ,  G01N15/147 ,  G01N21/031 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/85 ,  G01N2015/0294 ,  G01N2015/1006 ,  G01N2015/1472 ,  G01N2015/1486 ,  G02B7/28

    Abstract: An illuminiation system for increasing a light signal from an object passing through a reflection cavity. The reflection cavity is disposed between spaced-apart, opposed first and second surfaces disposed on opposite sides of a moving stream of objects. A light collection system is disposed substantially orthogonal to a plane passing through the surfaces and the stream so as to collect light that is scattered from or emitted by the objects as they pass through a field of view disposed between the first and second surfaces. A beam of light from a laser source is directed through the stream of moving objects in a direction nearly orthogonal to the stream (but slightly inclined) and lying in the plane that extends through the surfaces and the stream. Due to the reflection angle and the distance between the stream and the first surface, the point at which the light reflected from the first surface intersects the stream on a second pass is displaced from where it passed though the stream on its initial pass. The light is reflected back and forth between the surfaces a plurality of times, illuminating a different portion of the field of view with each pass until, having ranged over the field of view, the light exits the reflection cavity. The nullrecyclingnull of the light beam in this manner substantially improves the SNR of the detection system by increasing an average illumination intensity experienced by the objects in the stream.

    Abstract translation: 一种用于增加来自穿过反射腔的物体的光信号的照明系统。 反射腔设置在位于物体的移动物流的相对侧上的间隔开的相对的第一和第二表面之间。 光采集系统设置成基本上垂直于穿过表面和流的平面，以便收集当物体经过设置在第一表面和第二表面之间的视野时从物体散射或发射的光。 来自激光源的光束被引导通过几何垂直于流（但稍微倾斜）并且位于延伸穿过表面和流的平面中的移动物体流。 由于反射角度和流与第一表面之间的距离，从第一表面反射的光在第二遍上与流相交的点从其在其初始通过时通过流的位置偏移。 光在表面之间反复反射多次，每次通过照射视场的不同部分，直到在视场范围内，光离开反射腔。 以这种方式，光束的“回收”通过增加物流在物流中经历的平均照明强度，大大提高了检测系统的信噪比。

    Methods of calibrating an imaging system using calibration beads
    4.
    发明申请
    Methods of calibrating an imaging system using calibration beads 有权
    使用校准珠校准成像系统的方法

    公开(公告)号：US20030142289A1

    公开(公告)日：2003-07-31

    申请号：US10348193

    申请日：2003-01-16

    Applicant: Amnis Corporation

    Inventor： William E. Ortyn ,  David A. Basiji ,  Keith L. Frost ,  Brian E. Hall ,  Michael J. Seo

    IPC: G01P003/36

    CPC classification number: G01N15/1012 ,  G01N15/1459 ,  G01N15/1463 ,  G01N15/147 ,  G01N15/1475 ,  G01N2015/1018 ,  G01N2015/1025 ,  G01N2015/1075 ,  G01N2015/144 ,  G01N2015/1443 ,  G01N2015/1447 ,  G01N2015/1452 ,  G01N2015/1465 ,  G01N2015/1472 ,  G01N2015/1477 ,  G01N2015/1497 ,  G02B27/0012 ,  G06T7/20

    Abstract: When utilized in a flow imaging instrument, calibration beads provide a known data source that can be employed in various self-diagnostic, calibration and quality metric applications for the both the optical system of the flow imaging instrument, as well as the flow cell of the flow imaging instrument. Such data can be used to determine point spread functions associated with an imaging system, to determine a sensitivity of an imaging system, and to determine a focal point of the imaging system. Imagery collected from calibration beads can be used to determine core size and stability and TDI/flow speed synchronization. Calibration beads can be beneficially employed to enable stable system operation, even when very low sample concentration, or very small sample sizes are to be analyzed.

    Abstract translation: 当在流动成像仪器中使用时，校准珠粒提供已知的数据源，其可以用于流动成像仪器的光学系统的各种自我诊断，校准和质量度量应用，以及流量单元 流量成像仪。 这样的数据可用于确定与成像系统相关联的点扩散功能，以确定成像系统的灵敏度，并确定成像系统的焦点。 从校准珠收集的图像可用于确定核心尺寸和稳定性以及TDI /流速同步。 即使要分析非常低的样品浓度或非常小的样品量，也可以有利地使用校准珠来实现稳定的系统操作。

    Computational methods for the segmentation of images of objects from background in a flow imaging instrument
    5.
    发明申请
    Computational methods for the segmentation of images of objects from background in a flow imaging instrument 有权
    用于从流体成像仪器背景中分离物体图像的计算方法

    公开(公告)号：US20030086608A1

    公开(公告)日：2003-05-08

    申请号：US10200018

    申请日：2002-07-17

    Applicant: AMNIS CORPORATION

    Inventor： Keith L. Frost ,  James K. Riley

    IPC: G06K009/48

    CPC classification number: G02B27/0012 ,  G01N15/147 ,  G01N15/1475 ,  G01N21/6428 ,  G01N21/6456 ,  G01N21/6458 ,  G01N2015/1018 ,  G01N2015/1472 ,  G01N2015/1486 ,  G01N2015/1497

    Abstract: In automated computation-based interpretation of images, the accuracy and reliability of the detection and delineation of objects, known as nullobject segmentation,null is implemented so as to provide efficient performance. In a multi-step process, objects are first detected and captured into regions of interest (ROIs). Sets of pixels belonging to respective objects are then identified. Preferably object detection is achieved using both a two-dimensional (2D) low pass filter and a 2D edge enhancement filter. Two different gradient based edge enhancement filters are disclosed. One embodiment of the invention defines a (ROI) by first determining the center of objects by executing a plurality of decimations on the filtered image data, and then establishing object boundaries. In a second embodiment the ROI is defined by generating an amplitude histogram of the filtered image data, and for histograms exceeding a threshold determining by pixel which rows are to be included in the ROI.

    Abstract translation: 在图像的自动计算解释中，实现了被称为“对象分割”的对象的检测和描绘的准确性和可靠性，以提供有效的性能。 在多步骤过程中，首先检测对象并将其捕获到感兴趣的区域（ROI）中。 然后识别属于各个对象的像素集合。 优选地，使用二维（2D）低通滤波器和2D边缘增强滤波器来实现对象检测。 公开了两种不同的基于梯度的边缘增强滤波器。 本发明的一个实施例通过首先通过对经滤波的图像数据执行多个抽取，然后建立对象边界来首先确定对象的中心来定义（ROI）。 在第二实施例中，通过生成滤波图像数据的幅度直方图，并且对于直方图超过阈值来确定ROI将被包括在ROI中的哪个行来定义ROI。

    Method and apparatus for correcting crosstalk and spatial resolution for multichannel imaging
    6.
    发明申请
    Method and apparatus for correcting crosstalk and spatial resolution for multichannel imaging 有权
    用于校正多声道成像的串扰和空间分辨率的方法和装置

    公开(公告)号：US20030016882A1

    公开(公告)日：2003-01-23

    申请号：US10132059

    申请日：2002-04-24

    Applicant: Amnis Corporation is attached.

    Inventor： James K. Riley ,  Keith L. Frost

    IPC: G06T005/00 ,  G06T005/50 ,  G06K009/03 ,  G06K009/40 ,  G06K009/32

    CPC classification number: G06K9/00503 ,  G01J3/28 ,  G01J3/2803 ,  G01J3/32 ,  G01J3/36 ,  G01N15/147 ,  G01N15/1475 ,  G06K9/00127 ,  G06T5/50 ,  G06T7/32 ,  G06T7/33 ,  G06T2207/10036

    Abstract: A multichannel imaging system generates an ensemble of images for each field of view of an object. Each image in the ensemble is intended to contain information from only one source among a plurality of sources for the object. However, due to crosstalk, at least a portion of the signal from a first source appears in a channel intended for a second source. Because the accuracy of the correction will be degraded if the images in an ensemble are spatially misaligned with respect to one another, the spatial offset between images is determined and a correction is applied to substantially eliminate the offset. Then, a correction to the signals is determined to substantially reduce the contributions to the signal in a channel from the signals in other channels. The signal processing can be employed to process the output signals for each of a plurality of different disclosed imaging systems.

    Abstract translation: 多通道成像系统为对象的每个视场生成图像集合。 集合中的每个图像旨在包含来自对象的多个源中的一个源的信息。 然而，由于串扰，来自第一源的信号的至少一部分出现在用于第二源的通道中。 因为如果一个组合中的图像相对于彼此在空间上不对准，校正的精度将会降低，所以确定图像之间的空间偏移并且应用校正以基本上消除偏移。 然后，确定对信号的校正，以从其它通道中的信号基本上减少对信道中的信号的贡献。 信号处理可用于处理多个不同公开的成像系统中的每一个的输出信号。

    Auto focus for a flow imaging system
    7.
    发明申请
    Auto focus for a flow imaging system 有权
    流量成像系统的自动对焦

    公开(公告)号：US20040217256A1

    公开(公告)日：2004-11-04

    申请号：US10822170

    申请日：2004-04-09

    Applicant: Amnis Corporation

    Inventor： William E. Ortyn ,  Michael J. Seo ,  David A. Basiji ,  Keith L. Frost ,  David J. Perry

    IPC: G11B007/00 ,  G02B007/04

    CPC classification number: G06T7/20 ,  G01N15/1459 ,  G01N15/1463 ,  G01N2015/1018 ,  G01N2015/1443 ,  G01N2015/1447 ,  G01N2015/1452 ,  G01N2015/1497 ,  G02B27/0012 ,  G06K9/00134

    Abstract: A pair of optical gratings are used to modulate light from an object, and the modulated light from either optical is used to determine the velocity of the object. Each optical grating is offset from a reference focal point by the same distance, one grating being offset in a positive direction, the other in a negative direction. Signals produced in response to the modulated light can be processed to determine a direction in which a primary collection lens should be moved in order to improve a focus of the imaging system on the object. The lens is moved incrementally in the direction so determined, and the process is repeated until an optimal focus is achieved. In a preferred embodiment, the signals are weighted, so that the optical grating disposed closest to the optimal focus position contributes the most to velocity detection.

    Abstract translation: 一对光栅被用于调制来自物体的光，并且来自任一光学的调制光用于确定物体的速度。 每个光栅从参考焦点偏移相同的距离，一个光栅在正方向偏移，另一个光栅在负方向上偏移。 可以处理响应于调制光产生的信号以确定主收集透镜应该移动的方向，以便改善成像系统对物体的焦点。 透镜在如此确定的方向上逐渐移动，并重复该过程，直到达到最佳聚焦。 在优选实施例中，对信号进行加权，使得最靠近最佳聚焦位置布置的光栅最有助于速度检测。

    Multipass cavity for illumination and excitation of moving objects
    8.
    发明申请
    Multipass cavity for illumination and excitation of moving objects 有权
    多光腔用于照明和激发移动物体

    公开(公告)号：US20030137661A1

    公开(公告)日：2003-07-24

    申请号：US10355653

    申请日：2003-01-29

    Applicant: Amnis Corporation

    Inventor： William E. Ortyn ,  David A. Basiji ,  Keith L. Frost ,  N. Robert McRuer

    IPC: G01N021/00

    CPC classification number: G02B27/126 ,  G01J3/2803 ,  G01J3/2823 ,  G01N15/1434 ,  G01N15/147 ,  G01N21/031 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/85 ,  G01N2015/0294 ,  G01N2015/1006 ,  G01N2015/1472 ,  G01N2015/1486 ,  G02B7/28

    Abstract: An illumination system for increasing a light signal from an object passing through a reflection cavity. The reflection cavity is disposed between spaced-apart, opposed first and second surfaces disposed on opposite sides of a moving stream of objects. A light collection system is disposed substantially orthogonal to a plane passing through the surfaces and the stream so as to collect light that is scattered from or emitted by the objects as they pass through a field of view disposed between the first and second surfaces. A beam of light from a laser source is directed through the stream of moving objects in a direction nearly orthogonal to the stream (but slightly inclined) and lying in the plane that extends through the surfaces and the stream. Due to the reflection angle and the distance between the stream and the first surface, the point at which the light reflected from the first surface intersects the stream on a second pass is displaced from where it passed though the stream on its initial pass. The light is reflected back and forth between the surfaces a plurality of times, illuminating a different portion of the field of view with each pass until, having ranged over the field of view, the light exits the reflection cavity. The nullrecyclingnull of the light beam in this manner substantially improves the SNR of the detection system by increasing an average illumination intensity experienced by the objects in the stream.

    Abstract translation: 一种用于增加来自穿过反射腔的物体的光信号的照明系统。 反射腔设置在位于物体的移动物流的相对侧上的间隔开的相对的第一和第二表面之间。 光采集系统设置成基本上垂直于穿过表面和流的平面，以便收集当物体经过设置在第一表面和第二表面之间的视野时从物体散射或发射的光。 来自激光源的光束被引导通过几何垂直于流（但稍微倾斜）并且位于延伸穿过表面和流的平面中的移动物体流。 由于反射角度和流与第一表面之间的距离，从第一表面反射的光在第二遍上与流相交的点从其在其初始通过时通过流的位置偏移。 光在表面之间反复反射多次，每次通过照射视场的不同部分，直到在视场范围内，光离开反射腔。 以这种方式，光束的“回收”通过增加物流在物流中经历的平均照明强度，大大提高了检测系统的信噪比。

    Spectral deconvolution of fluorescent markers
    9.
    发明申请
    Spectral deconvolution of fluorescent markers 有权
    荧光标记的光谱去卷积

    公开(公告)号：US20030020908A1

    公开(公告)日：2003-01-30

    申请号：US10175543

    申请日：2002-06-18

    Applicant: Amnis Corporation

    Inventor： Keith L. Frost ,  James K. Riley ,  David A. Basiji ,  William E. Ortyn

    IPC: G01N021/64

    CPC classification number: G01J3/10 ,  G01J3/14 ,  G01J3/28 ,  G01J3/4406 ,  G01N15/147 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6456 ,  G01N2015/1472 ,  G01N2015/1486 ,  G01N2015/1497 ,  G01N2021/6423 ,  G01N2021/6441 ,  G01N2201/1293

    Abstract: In an instrument generating images from the fluorescent emissions of a plurality of fluorescent dyes carried by objects in a flow stream, spectral dispersion is used to expand the images of the objects along one axis of a two-dimensional photosensor array according to emission wavelength. The dispersion is unable to completely separate the emissions of a plurality of dyes because the emission spectra of the dyes overlap in wavelength. The method of the present invention accomplishes accurate estimation of the intensity of the light received from each of a plurality of dyes by decomposing the two dimensional spectrally dispersed images into a set of characteristic parameters using either an iterative curve fitting optimization method or a linear algebraic method.

    Abstract translation: 在从由流体中的物体携带的多个荧光染料的荧光发射产生图像的仪器中，使用光谱色散来根据发射波长来扩展沿着二维光电传感器阵列的一个轴的物体的图像。 由于染料的发射光谱在波长上重叠，所以色散不能完全分离多种染料的发射。 本发明的方法通过使用迭代曲线拟合优化方法或线性代数方法将二维光谱分散图像分解成一组特征参数，从而准确估计从多种染料中的每一种接收的光的强度 。

    Method and apparatus for reading reporter labeled beads
    10.
    发明申请
    Method and apparatus for reading reporter labeled beads 失效
    阅读报道标记珠粒的方法和装置

    公开(公告)号：US20020094116A1

    公开(公告)日：2002-07-18

    申请号：US09976237

    申请日：2001-10-12

    Applicant: Amnis Corporation

    Inventor： Keith L. Frost ,  David A. Basiji ,  Richard A. Bauer ,  Rosalynde J. Finch ,  William E. Ortyn ,  David J. Perry

    IPC: G06K009/00

    CPC classification number: G02B27/148 ,  C12Q1/6816 ,  C40B20/04 ,  C40B30/04 ,  C40B40/08 ,  C40B60/10 ,  G01N15/147 ,  G01N15/1475 ,  G01N2015/1447 ,  G01N2015/1472 ,  G01N2015/1479 ,  G01N2015/1497 ,  G02B7/28 ,  G02B27/0012 ,  G02B27/1013 ,  G02B27/126 ,  G02B27/144 ,  G02B27/145 ,  C12Q2563/179 ,  C12Q2563/155 ,  C12Q2563/107

    Abstract: Combinatorially-synthesized deoxyribonucleic acid (DNA) oligonucleotides attached to encoded beads that are hybridized to amplified and labeled genomic DNA or ribonucleic acid (RNA) are analyzed using a flow imaging system. Oligonucleotides and corresponding reporters are bound to the surfaces of a plurality of small beads such that different beads bear different oligo sequences. Each bead bears a unique optical signature comprising a predefined number of unique reporters, where each reporter comprises a predefined combination of different fluorochromes. The composite spectral signature in turn identifies the unique nucleotide sequence of its attached oligo chains. This optical signature is rapidly decoded using an imaging system to discriminate the different reporters attached to each bead in a flow in regard to color and spatial position on the bead.

    Abstract translation: 使用流动成像系统分析与编码的珠偶联的与扩增和标记的基因组DNA或核糖核酸（RNA）杂交的组合合成的脱氧核糖核酸（DNA）寡核苷酸。 寡核苷酸和相应的报道结合到多个小珠的表面，使得不同的珠具有不同的寡核苷酸序列。 每个珠具有唯一的光学特征，其包含预定数量的唯一记录器，其中每个报告者包含不同荧光染料的预定组合。 复合光谱特征依次识别其连接的寡链的唯一核苷酸序列。 使用成像系统快速解码该光学特征，以便鉴别珠子上颜色和空间位置的流动中附着于每个珠粒的不同报告者。

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