Zhang Nicholas, Fang Zhou, Kadakia Priyam, Guo Jamie, Vijay Dakshin, Thapa Manoj, Dembowitz Samuel, Grakoui Arash, Coskun Ahmet F
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
Interdisciplinary Bioengineering Graduate Program, Georgia Institute of Technology, Atlanta, GA, USA.
Lab Chip. 2025 Sep 12. doi: 10.1039/d5lc00286a.
Spatial omics technologies have revolutionized the field of biology by enabling the visualization of biomolecules within their native tissue context. However, the high costs associated with proprietary instrumentation, specialized reagents, and complex workflows have limited the broad application of these techniques. In this study, we introduce Python-based robotic imaging and staining for modular spatial omics (PRISMS), an open-sourced, automated multiplexing pipeline compatible with several biospecimen targets and streamlined microscopy software tools. PRISMS utilizes a liquid handling robot with thermal control to enable the rapid and automated staining of RNA and protein samples. The modular sample holders and Python control facilitate high-throughput, single-molecule fluorescence imaging on widefield and confocal microscopes. We successfully demonstrated the versatility of PRISMS by imaging tissue slides and adherent cells. We demonstrate that PRISMS can be utilized to perform super-resolved imaging, such as super-resolution radial fluctuations (SRRF). PRISMS is a powerful tool that can be used to democratize spatial omics by providing researchers with an accessible, reproducible, and cost-effective solution for multiplex imaging. Specifically, PRISMS is an open-source, automated multiplexing pipeline for spatial omics, compatible with several sample types and Nikon NIS Elements Basic Research software, as well as Python-based biodevices. It performs high-throughput, single-molecule fluorescence imaging both on widefield and confocal microscopes, and can be used to perform super-resolved imaging, such as SRRF. Overall, PRISMS is a powerful tool that can be used to democratize spatial omics by providing researchers with an accessible, reproducible, and cost-effective solution for multiplex imaging. This open-source platform will enable researchers to push the boundaries of spatial biology and make groundbreaking discoveries.
空间组学技术通过在天然组织环境中实现生物分子的可视化，给生物学领域带来了变革。然而，与专有仪器、专用试剂和复杂工作流程相关的高成本限制了这些技术的广泛应用。在本研究中，我们介绍了基于Python的模块化空间组学机器人成像与染色技术（PRISMS），这是一种开源的自动化多重分析流程，与多种生物样本靶点兼容，并集成了简化的显微镜软件工具。PRISMS利用一台带有温度控制的液体处理机器人，实现RNA和蛋白质样本的快速自动化染色。模块化样本架和Python控制功能有助于在宽视场显微镜和共聚焦显微镜上进行高通量单分子荧光成像。我们通过对组织切片和贴壁细胞成像，成功展示了PRISMS的多功能性。我们证明PRISMS可用于进行超分辨成像，如超分辨率径向涨落（SRRF）。PRISMS是一个强大的工具，通过为研究人员提供一种可及、可重复且经济高效的多重成像解决方案，可推动空间组学的普及。具体而言，PRISMS是一个用于空间组学的开源自动化多重分析流程，与多种样本类型以及尼康NIS Elements基础研究软件和基于Python的生物设备兼容。它能在宽视场显微镜和共聚焦显微镜上进行高通量单分子荧光成像，还可用于进行超分辨成像，如SRRF。总体而言，PRISMS是一个强大的工具，通过为研究人员提供一种可及、可重复且经济高效的多重成像解决方案，可推动空间组学的普及。这个开源平台将使研究人员能够突破空间生物学的界限，做出开创性的发现。
