Soft tissue sarcoma (STS), a rare and diverse cancer in soft tissues, often leads to metastases despite optimal treatment. Lacking information on the exact role of essential metals in STS progression highlights the need for development of methods enabling metal quantification at the single cell level across different STS stages. Very few analytical techniques are capable of quantifying metals at single cell, or even sub-cellular, resolution. Analysis of individual cells in suspension via single-cell ICP-MS enables rapid data acquisition, but this approach has limitations, including low transport efficiency and lack of information on the cell’s environment. This project aims to develop innovative single cell laser ablation-ICP-MS (LA-ICP-MS) methods to explore the intricate relationship between essential metals and STS biochemistry. The project involves in-depth methodological development for high-efficiency, high-resolution single cell LA-ICP-MS analysis, also introducing automated LA sampling software. It will allow a better understanding of STS progression by unveiling metal homeostasis at the cellular level, revealing subcellular distribution, and unraveling cellular interactions in cell culture samples. Subsequently, multiplexed high-resolution LA-ICP-MS will be developed to map metal and protein (via tagging methods) distributions in STS-patient-derived xenograft (STS-PDX) tissue samples at various stages, and findings will be validated against real patient samples.