CIL:45850, 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学谱分析检测法，该方法可在不损害高通量提取丰富定量特征谱能力的前提下，用尽可能多的荧光标记物为细胞“上色”。该检测法可识别七种主要细胞组分。在一项生物活性化合物的先导筛选中，该检测法可检测到一系列细胞表型，并基于细胞学谱将具有相似注释蛋白靶点或化学结构的化合物进行聚类。研究结果表明，该检测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，也能检测到其生物学效应。这种基于图像的检测法提供了一种无偏策略，可用于表征化合物及疾病相关的细胞状态，为未来的探针发现研究提供支撑。

借助细胞绘图（cell-painting）检测法，布罗德研究所（Broad Institute）构建了一套经约30000种化合物处理的U2OS骨肉瘤细胞的特征谱参考数据集。该化合物集包含20000种DOS衍生化合物，以及通过PubChem分析筛选得到的10000种化学结构多样、生物学活性谱广泛的MLI化合物，另有2500种用作参照的已知生物活性化合物。其中，DOS衍生化合物结构多样且立体化学特征丰富，其结构与当前的MLSMR化合物存在显著差异。该化合物集还包含MLPCN中心从相关项目中提名的267种独特化合物，这些项目旨在识别具有相似活性的新型化学系列。

该实验包含413块微孔板，每块板含384个孔，每个孔包含9个视野。每个视野通过五个通道（检测波长）成像，每个通道以独立的灰度16位TIFF图像文件格式存储。