A high-throughput technique to map cell images to cell positions using a 3D imaging flow cytometer.
A high-throughput technique to map cell images to cell positions using a 3D imaging flow cytometer.
复制标题
使用3D成像流式细胞仪将细胞图像映射到细胞位置的高通量技术。
DOI：
10.1073/pnas.2118068119
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发表时间：
2022-02-22
影响因子：
11.1
通讯作者：
Lo YH
中科院分区：
文献类型：
--
作者：
Zhang Z;Tang R;Chen X;Waller L;Kau A;Fung AA;Gutierrez B;An C;Cho SH;Shi L;Lo YH
本文演示了一种高通量的技术，将细胞图像映射到细胞位置。该技术使用三维（3D）成像流式细胞仪以1，000个细胞/s的通量记录多参数3D细胞图像，并使用细胞放置机器人以先进先出的方式将来自成像系统的退出细胞放置在滤板上，因此板上的细胞具有与成像细胞相同的顺序。开发了创新的算法来匹配来自成像和放置模块的细胞序列，以检测和消除错误，以确保高精度。该技术在单细胞分子分析和单细胞图像分析之间形成了前所未有的桥梁，将表型和基因型分析与单细胞分辨率连接起来。我们开发了一种高通量技术，将单个细胞的位置与其具有单细胞分辨率的三维（3D）成像特征联系起来。该技术特别适用于非贴壁细胞，其中现有的空间生物学方法将细胞特性与其在实体组织中的位置相关联并不适用。我们的设计包括两个部分，如下：使用定制的3D成像流式细胞仪（3D-IFC）以高通量（500至1，000个细胞/s）记录3D细胞图像，并使用机器人细胞放置平台（CPP）以先进先出（FIFO）方式分配细胞。为了防止由于违反FIFO原则而导致的错误，我们发明了一种使用标记珠和DNA测序软件来检测错误的方法。人类癌细胞系的实验证明了在不到10分钟的时间内将细胞的3D侧向散射和荧光图像以及二维（2D）透射图像映射到其在膜过滤器上的位置的可行性，大约100，000个细胞。虽然目前的工作使用我们专门设计的3D成像流式细胞仪来产生3D细胞图像，我们的方法可以支持其他成像模式。该技术和方法在单细胞图像分析和单细胞分子分析之间架起了一座桥梁。
This article demonstrates a high-throughput technique to map cell images to cell positions. The technology uses a three-dimensional (3D) imaging flow cytometer to record multiparameter 3D cell images at a throughput of 1,000 cells/s and a cell placement robot to place the exiting cells from the imaging system on a filter plate in a first-in–first-out manner so the cells on the plate have the same order as the cells that are imaged. Innovative algorithms were developed to match the cell sequences from the imaging and placement modules to detect and eliminate errors to ensure high accuracy. The technology forms an unprecedented bridge between single-cell molecular analysis and single-cell image analysis to connect phenotype and genotype analysis with single-cell resolution. We develop a high-throughput technique to relate positions of individual cells to their three-dimensional (3D) imaging features with single-cell resolution. The technique is particularly suitable for nonadherent cells where existing spatial biology methodologies relating cell properties to their positions in a solid tissue do not apply. Our design consists of two parts, as follows: recording 3D cell images at high throughput (500 to 1,000 cells/s) using a custom 3D imaging flow cytometer (3D-IFC) and dispensing cells in a first-in–first-out (FIFO) manner using a robotic cell placement platform (CPP). To prevent errors due to violations of the FIFO principle, we invented a method that uses marker beads and DNA sequencing software to detect errors. Experiments with human cancer cell lines demonstrate the feasibility of mapping 3D side scattering and fluorescent images, as well as two-dimensional (2D) transmission images of cells to their locations on the membrane filter for around 100,000 cells in less than 10 min. While the current work uses our specially designed 3D imaging flow cytometer to produce 3D cell images, our methodology can support other imaging modalities. The technology and method form a bridge between single-cell image analysis and single-cell molecular analysis.
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