Simultaneous visualization of the relationship between multiple biomolecules and their ligands or small molecules at the nanometer scale in cells will enable greater understanding of how biological processes operate. We present here high-definition multiplex ion beam imaging (HD-MIBI), a secondary ion mass spectrometry approach capable of high-parameter imaging in 3D of targeted biological entities and exogenously added structurally-unmodified small molecules. With this technology, the atomic constituents of the biomolecules themselves can be used in our system as the “tag” and we demonstrate measurements down to ~30 nm lateral resolution. We correlated the subcellular localization of the chemotherapy drug cisplatin simultaneously with five subnuclear structures. Cisplatin was preferentially enriched in nuclear speckles and excluded from closed-chromatin regions, indicative of a role for cisplatin in active regions of chromatin. Unexpectedly, cells surviving multi-drug treatment with cisplatin and the BET inhibitor JQ1 demonstrated near total cisplatin exclusion from the nucleus, suggesting that selective subcellular drug relocalization may modulate resistance to this important chemotherapeutic treatment. Multiplexed high-resolution imaging techniques, such as HD-MIBI, will enable studies of biomolecules and drug distributions in biologically relevant subcellular microenvironments by visualizing the processes themselves in concert, rather than inferring mechanism through surrogate analyses.

同时可视化多个生物分子与其配体或细胞中纳米级小分子之间的关系，将有助于更好地了解生物过程的运作方式。我们在此展示了高清多重离子束成像 (HD-MIBI)，这是一种二次离子质谱法，能够对目标生物实体和外源添加的结构未修饰小分子进行 3D 高参数成像。使用这项技术，生物分子本身的原子成分可以在我们的系统中用作“标签”，我们展示了低至 ~30 nm 横向分辨率的测量。我们将化疗药物顺铂的亚细胞定位同时与五个亚核结构相关联。顺铂优先富集在核斑点中，并被排除在封闭染色质区域之外，表明顺铂在染色质活性区域中的作用。出乎意料的是，在用顺铂和 BET 抑制剂 JQ1 进行多药治疗后幸存下来的细胞表现出几乎完全从细胞核中排除顺铂，这表明选择性亚细胞药物重新定位可能会调节对这种重要化疗治疗的抵抗力。多路高分辨率成像技术，如 HD-MIBI，将通过协同可视化过程本身，而不是通过替代分析来推断机制，从而能够研究生物相关亚细胞微环境中的生物分子和药物分布。用顺铂和 BET 抑制剂 JQ1 进行多药治疗后存活下来的细胞表明，顺铂几乎完全被排除在细胞核之外，这表明选择性亚细胞药物重新定位可能会调节对这种重要化疗治疗的抵抗力。多路高分辨率成像技术，如 HD-MIBI，将通过协同可视化过程本身，而不是通过替代分析来推断机制，从而能够研究生物相关亚细胞微环境中的生物分子和药物分布。用顺铂和 BET 抑制剂 JQ1 进行多药治疗后存活下来的细胞表明，顺铂几乎完全被排除在细胞核之外，这表明选择性亚细胞药物重新定位可能会调节对这种重要化疗治疗的抵抗力。多路高分辨率成像技术，如 HD-MIBI，将通过协同可视化过程本身，而不是通过替代分析来推断机制，从而能够研究生物相关亚细胞微环境中的生物分子和药物分布。