Abstract

Spatial proteomics offers critical insights into tissue heterogeneity but remains technically challenging, particularly for archived clinical samples and analyses at single-cell or subcellular resolution. In this webinar, we introduce filter-aided expansion proteomics (FAXP), a hydrogel-based tissue expansion strategy enabling high-resolution, mass spectrometry–based spatial proteomics of formalin-fixed paraffin-embedded (FFPE) tissues.

FAXP integrates filter-aided in-gel digestion within customized C18 tip devices, improving workflow stability, throughput, and reproducibility. Compared to the original ProteomEx method, FAXP delivers a 14.5-fold improvement in volumetric resolution, over 8-fold higher peptide yield, a 255% increase in protein identification, and a 50% reduction in sample preparation time.

Compatible with archived FFPE tissues, FAXP can be combined with laser capture microdissection to enable image-guided proteomic analysis at single-cell and subcellular resolution. This webinar will outline the FAXP workflow and highlight its applications in high-resolution spatial proteomics, including studies of human colorectal cancer.

 Highlights

- Introduction to filter-aided expansion proteomics (FAXP).

- High-resolution spatial proteomics using FAXP combined with LCM.

- Application to archived FFPE tissues.

- Opportunities and limitations.