Imaging with high chemical and spatial resolutions using mass spectrometry

The National Institute of General Medical Sciences awarded the University of Florida $1,892,969 on September 1, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to develop novel mass spectrometry technologies for high-resolution molecular imaging of tissue samples.

The research addresses critical deficiencies in imaging mass spectrometry (IMS) by advancing chemistries and next-generation analytical techniques that improve both chemical differentiation and spatial resolution. Current commercial IMS platforms are limited to approximately 20 micrometers spatial resolution and struggle to distinguish overlapping molecular masses in tissue samples, which distorts the interpretation of biochemical maps. The funded work will develop technologies enabling sub-cellular resolution imaging and better structural identification of biomolecules, with applications to understanding the molecular basis of health and disease and supporting targeted diagnostics and therapeutics.

Work is performed in Gainesville, Florida, with performance continuing through August 31, 2030. The University of Florida Division of Sponsored Research administers the award as prime recipient.

Abstract

Molecular studies in targeted diagnostics and therapeutics have historically been limited to systems with some a priori knowledge of the specific molecules or biological pathways to be studied. New technologies have allowed for more comprehensive analyses of biological and clinical samples in recent years, which are in turn enabling greater insights into basic cellular biology and new opportunities for curative therapies. However, critical to realizing this potential is the continual development of analytical technologies and their application, in collaboration with biologists and clinicians, to important problems in human health. The overarching theme of my research program is to develop novel chemistries and next-generation analytical mass spectrometry (MS) technologies to better understand the molecular basis of health and disease. The use of mass spectrometry as a molecular imaging modality combines the high level of molecular specificity provided by the mass spectrometer with the spatial fidelity of a microscopic imaging approach. By this, imaging mass spectrometry (IMS) provides for the untargeted mapping of a wide array of biomolecules in tissue specimens. However, severe deficiencies remain in the differentiation and structural identification of molecules detected during IMS experiments due to the enormous chemical complexity of tissue samples. Additionally, despite the success and growing availability of IMS, spatial resolution is limited to ~20 ΜM on most commercial platforms and sub-cellular resolutions are generally not accessible. The failure to adequately resolve compounds and structures results in ION images consisting of multiple different compounds with overlapping masses and precludes resolution of sub-cellular features. These distorted pictures of molecular distributions cloud the interpretation of biochemical maps produced by IMS and prevents complete and accurate understanding of cellular compositions and functions. We will follow two broad, distinct directions to address these limitations, focusing on novel instrumentation, gas- phase reactions, sample preparation strategies, and multimodal bioinformatics workflows for a range of tissues and analytes. In the first direction, we will develop mass spectrometry-compatible tissue expansion protocols that physically magnify the tissue substrate, resulting in higher effective spatial resolutions compared to imaging of UN-magnified tissues with the same diameter laser beam. Expansion imaging mass spectrometry (exims) will be complemented with computational "upsampling" via image fusion to enable high spatial resolution (< 2 ΜM) multiplexed IMS to be performed on existing commercial platforms. In the second direction, we will develop instrumentation and novel gas-phase ION/ION reactions that allow for unique identification of metabolites, lipids, and proteins. These reactions offer rapid and flexible means for molecular transformations without manipulating the tissue sample and can result in improved detection limits and more extensive chemical structural information. Accurate identification of biochemical pathways altered during development and dysfunction is key to designing novel treatment strategies in areas such as diabetes, infectious disease, neurodegeneration, and cancer.