CIL:45809, Homo sapiens. In Cell Image Library

Gustafsdottir et al. (doi:10.1371/journal.pone.0080999) have developed a multiplex cytological profiling assay that "paints the cell" with as many fluorescent markers as possible without compromising our ability to extract rich, quantitative profiles in high throughput. The assay detects seven major cellular components. In a pilot screen of bioactive compounds, the assay detected a range of cellular phenotypes and it clustered compounds with similar annotated protein targets or chemical structure based on cytological profiles. The results demonstrate that the assay captures subtle patterns in the combination of morphological labels, thereby detecting the effects of chemical compounds even though their targets are not stained directly. This image-based assay provides an unbiased approach to characterize compound- and disease-associated cell states to support future probe discovery.

Using the cell-painting assay, the Broad Institute has assembled a reference dataset of profiles for U2OS osteosarcoma cells treated with ~30,000 compounds. The compound collection includes DOS-derived compounds (20,000), as well as chemically diverse MLI compounds with biologically diverse performance identified through analysis of PubChem (10,000), and known bioactive compounds to serve as landmarks (2,500). The DOS compounds consist of structurally diverse and stereochemically rich compounds with structures distinct from the current MLSMR. The compound collection also includes 267 distinct compounds nominated by MLPCN Centers from projects for which the Centers would like to identify new chemical series with similar activities.

The experiment consists of 413 microtiter plates. Each plate has 384 wells. Each well has 9 fields of view. Each field was imaged in five channels (detection wavelengths), and each channel is stored as a separate, grayscale 16-bit TIFF image file.

Gustafsdottir等人（doi:10.1371/journal.pone.0080999）开发了一种多重细胞形态谱分析检测法（multiplex cytological profiling assay），该方法可使用尽可能多的荧光标记物为细胞进行染色标记，同时不影响我们以高通量方式获取丰富的定量细胞谱的能力。该检测方法可识别七种主要的细胞组分。在针对生物活性化合物的初步筛选中，该检测方法可捕获多种细胞表型，并能基于细胞谱对带有相似注释蛋白靶点或相似化学结构的化合物进行聚类分析。研究结果表明，该检测方法可捕捉形态学标记组合中的细微特征，即便化合物的靶点未被直接染色，也能识别其对细胞产生的影响。这种基于成像的检测方法为表征化合物及疾病相关细胞状态提供了无偏倚的研究路径，可为未来的探针开发提供支撑。

借助细胞绘画检测法（Cell Painting assay），布罗德研究所针对经约30000种化合物处理的U2OS骨肉瘤细胞，构建了一套细胞谱参考数据集。该化合物库包含：源自DOS的化合物（20000种）、通过PubChem分析筛选得到的化学结构多样且生物学活性各异的MLI化合物（10000种），以及作为参照标杆的已知生物活性化合物（2500种）。DOS类化合物具有结构多样性高、立体化学丰富的特点，其结构与当前的MLSMR库存在显著差异。该化合物库还包含由MLPCN中心从相关项目中提名的267种独特化合物，这些项目旨在筛选出具有相似活性的新型化学序列。

该实验共涉及413块微孔板。每块板包含384个孔，每个孔设有9个视场。每个视场通过5个通道（检测波长）进行成像，每个通道的图像均以独立的灰度16位TIFF文件格式存储。