Combining padlock probe-based RNA detection with protein and DNA readouts into a flexible, fully controllable spatial omics methodology (Padlock Plus ‘X’)
The human body is a complex system made up of many different types of cells, each with its own unique role. These cells must work together in harmony to keep us healthy. When this balance is disrupted, it can lead to diseases, including cancer. To fully understand how our cells interact and function, scientists need to study them not only individually but also in the context of their surroundings — this is called "spatial biology."
Over the last decade, tools and technologies for spatial biology have advanced significantly, helping researchers gain valuable insights into how cells behave in health and disease. These tools allow scientists to explore the locations and interactions of biomolecules like DNA, RNA, and proteins within their cellular environment. This approach has already improved our understanding of cancer and human development.
In our project, we aimed to develop a protocol to study selected proteins, RNA, and DNA all within the same human tissue sample. This would provide a more comprehensive view of cellular processes. To start, we optimized a method to visualize RNA and protein together within cells. This achievement is a critical step toward creating a powerful tool for studying human biology in its full complexity.
Fluorescence microscopy images of HUVEC cells (30 min stimulation with TNF-alpha) displaying the combinatorial approach between In Situ Sequencing (ISS) for intronic RNA detection (CXCL1-2-8) and Immuno-SABER for protein detection (SON, H2A1, TFAM, Vimentin). Dotted line is used to display the cell nuclei.