Maximising Spatial Biology

Spatial biology, the study of the spatial organization of cells and molecules within tissue, is revolutionizing our understanding of complex biological processes. Traditional methods often provide limited insights due to their focus on thin tissue sections. However, a groundbreaking workflow combining 3D imaging with multiplex spatial analysis is changing the game.

This innovative approach allows for the comprehensive examination of large tissue samples while retaining the ability to analyze specific regions in depth, all from the same specimen. In a proof of concept study, we utilized 3D-immunofluorescence (3D-IF) staining and tissue clearing to prepare mouse brain hemispheres for 3D light sheet imaging with the UltraMicroscope Blaze™. With the full specimen’s 3D view, we were able to identify target regions in order to prepare tissue sections that precisely cover these specific parts of the specimen, a process termed light sheet guided histology.

The tissue sections were further analyzed with MACSima™ Imaging Cyclic Staining (MICS), providing expression levels of up to hundreds of protein markers from individual cells on a single sample. Remarkably, the fluorochrome conjugates used for 3D-IF staining remained detectable after sectioning, enabling us to verify the location of our target region and markers. Furthermore, the epitopes remained stable throughout the entire process of sample preparation for 3D imaging, 3D imaging itself and sample preparation for MICS. Thus, we have demonstrated that it is possible to apply our MICS technology to a previously cleared tissue.

The workflow, termed light sheet guided spatial biology, was performed using the UltraMicroscope Blaze for 3D imaging, and the MACSima Imaging System for spatial biology from Miltenyi Biotec. By combining 3D imaging with multiplex analysis, this workflow offers a comprehensive understanding of biological structures without sacrificing finer details. This approach is crucial for unraveling the complexities of cellular and molecular processes, advancing drug-based treatment methods, and enhancing diagnostic capabilities.

Figure 1: Illustration of a spatial biology workflow that combines MICS technology with prior 3D imaging.

Moreover, this technology holds immense potential for applications such as identifying metastasis in large tissues, assessing intratumor infiltration of therapeutic cells, and understanding neurodegenerative disorders. As the workflow continues to evolve, with ongoing research focusing on diverse tissues and applications, its impact on spatial biology will undoubtedly grow, paving the way for new discoveries and therapeutic interventions.

To read the full study, visit https://tinyurl.com/2s5sd3zv or scan the QR code.