Abstract
Biological tissues exhibit complex spatial heterogeneity that directs the functions of multicellular organisms. Quantifying protein expression is essential for elucidating processes within complex biological assemblies. Imaging mass spectrometry (IMS) is a powerful emerging tool for mapping the spatial distribution of metabolites and lipids across tissue surfaces, but technical challenges have limited the application of IMS to the analysis of proteomes. Methods for probing the spatial distribution of the proteome have generally relied on the use of labels and/or antibodies, which limits multiplexing and requires a priori knowledge of protein targets. Past efforts to make spatially resolved proteome measurements across tissues have had limited spatial resolution and proteome coverage and have relied on manual workflows. Here, we demonstrate an automated approach to imaging that utilizes label-free nanoproteomics to analyze tissue voxels, generating quantitative cell-type-specific images for >2000 proteins with 100-µm spatial resolution across mouse uterine tissue sections preparing for blastocyst implantation.
Highlights
Biological tissues exhibit complex spatial heterogeneity that directs the functions of multicellular organisms
Proteomic approaches have been combined with isolation techniques such as laser capture microdissection (LCM) and fluorescence-activated cell sorting (FACS); these applications are narrow to date, due to the limited amount of sample mass obtainable[8,20,21,22,23,24]
Our approach combines existing technology with a suite of technologies recently developed in our lab to achieve the robustness, sensitivity, and throughput that are essential for proteome-level imaging
Summary
Biological tissues exhibit complex spatial heterogeneity that directs the functions of multicellular organisms. Imaging mass spectrometry (IMS) is a powerful emerging tool for mapping the spatial distribution of metabolites and lipids across tissue surfaces, but technical challenges have limited the application of IMS to the analysis of proteomes. In an IMS experiment, a probe, which may be a laser, ion beam, or liquid junction, is rastered across a surface to desorb or extract biomolecules that are directly analyzed by mass spectrometry (MS). This allows for the creation of detailed spatial maps that reveal the native distribution of biomolecules at the surface without labels or pretreatment. There is currently no IMS technology capable of in-depth proteome imaging
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