This research is aimed to push the boundaries of imaging mass spectrometry with unique biological applications directed toward the chemical characterization of single biological cells. A special emphasis is placed upon probing the lipid distributions and the role of small molecule neurotransmitters embedded in neurotransmitter vesicles primed for exocytosis and brain lipids in Alzheimer's disease. Model systems include liposome cell mimics, mast cells, PC12 cells and brain tissue where membrane structure is changing. The central approach is to utilize an advanced bio-analytical mass spectrometry-based protocol to acquire two and three dimensional molecule-specific image information at the nano-scale level. Transformational mass spectrometry instrumentation for secondary ion mass spectrometry (SIMS) is proposed to complete this mission. Special emphasis is placed upon implementing a novel gas cluster ion beam (GCIB), consisting of an Ar4000+ projectile, which has been shown to desorb molecules without breaking bonds and without chemical damage buildup on the sample. There are four specific aims for this proposal. First, we propose to combine 3- dimensional imaging with a tightly focused C60+ ion source and a GCIB source especially tuned by chemistry to dramatically enhance spatial resolution and ionization efficiency in imaging. This will increase sensitivity and therefore spatial resolution as we push t the limits obtained by dynamic SIMS. Second, the effectiveness of these protocols will be verified using well-characterized liposome particles as stand-ins for biological cells. Third, developments in the first two aims will be implemented to image the substructure of vesicles in single cells in order to gain a better understanding of what regulates chemical communication via exocytosis. These studies will provide fundamentally important information about how neurotransmitter release is regulated for plasticity and pharmacology. Fourth, metabolic labeling with stable isotope-encoded lipid and protein species will be used to elucidate spatial and temporal changes in the molecular architecture of single nerve cells. We will then use this imaging methodology to study protein aggregation in single nerve cells and a model for Alzheimer's disease. The overarching objective is to establish the unique imaging strategy proposed here as a valuable new tool for use by the larger biological community.

Public Health Relevance

We propose to make significant advances in imaging mass spectrometry in three dimensions which are needed to test dynamical theories of biological function at the single cell level. Special emphasis is placed upon probing the lipid distributions and the role of small molecule neurotransmitters embedded in fused vesicles which might play a role in influencing, for example, Alzheimer's disease states. An overarching objective is to provide a unique capability to the larger biological and public health research community.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
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Sheeley, Douglas
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Pennsylvania State University
Schools of Arts and Sciences
University Park
United States
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Shen, Kan; Tarolli, Jay G; Winograd, Nicholas (2016) Cluster secondary ion mass spectrometry imaging of interfacial reactions of TiO2 microspheres embedded in ionic liquids. Rapid Commun Mass Spectrom 30:379-85
Tian, Hua; Wucher, Andreas; Winograd, Nicholas (2016) Dynamic Reactive Ionization with Cluster Secondary Ion Mass Spectrometry. J Am Soc Mass Spectrom 27:285-92
Tian, Hua; Wucher, Andreas; Winograd, Nicholas (2016) Reduce the matrix effect in biological tissue imaging using dynamic reactive ionization and gas cluster ion beams. Biointerphases 11:02A320
Tian, Hua; Maciążek, Dawid; Postawa, Zbigniew et al. (2016) CO2 Cluster Ion Beam, an Alternative Projectile for Secondary Ion Mass Spectrometry. J Am Soc Mass Spectrom 27:1476-82
Tarolli, Jay Gage; Bloom, Anna; Winograd, Nicholas (2016) Multimodal image fusion with SIMS: Preprocessing with image registration. Biointerphases 11:02A311
Bloom, Anna N; Tian, Hua; Winograd, Nicholas (2015) C60-SIMS imaging of nanoparticles within mammalian cells. Biointerphases 11:02A306
Majdi, Soodabeh; Berglund, E Carina; Dunevall, Johan et al. (2015) Electrochemical Measurements of Optogenetically Stimulated Quantal Amine Release from Single Nerve Cell Varicosities in Drosophila Larvae. Angew Chem Int Ed Engl 54:13609-12
Li, Xianchan; Majdi, Soodabeh; Dunevall, Johan et al. (2015) Quantitative measurement of transmitters in individual vesicles in the cytoplasm of single cells with nanotip electrodes. Angew Chem Int Ed Engl 54:11978-82
Winograd, Nicholas (2015) Imaging mass spectrometry on the nanoscale with cluster ion beams. Anal Chem 87:328-33
Bloom, Anna; Winograd, Nicholas (2014) Dye-Enhanced imaging of mammalian cells with SIMS. Surf Interface Anal 46:177-180

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