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)
Project #
Application #
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Sheeley, Douglas
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Pennsylvania State University
Schools of Arts and Sciences
University Park
United States
Zip Code
Sämfors, Sanna; Ewing, Andrew G; Fletcher, John S (2018) Benefits of NaCl addition for ToF-SIMS analysis including the discrimination of diacylglyceride and triacylglyceride ions. Rapid Commun Mass Spectrom :
Tian, Hua; Six, David A; Krucker, Thomas et al. (2017) Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C60-Secondary Ion Mass Spectrometry. Anal Chem 89:5050-5057
Dunevall, Johan; Majdi, Soodabeh; Larsson, Anna et al. (2017) Vesicle impact electrochemical cytometry compared to amperometric exocytosis measurements. Curr Opin Electrochem 5:85-91
Ren, Lin; Pour, Masoumeh Dowlatshahi; Majdi, Soodabeh et al. (2017) Zinc Regulates Chemical-Transmitter Storage in Nanometer Vesicles and Exocytosis Dynamics as Measured by Amperometry. Angew Chem Int Ed Engl 56:4970-4975
Tian, Hua; Sparvero, Louis J; Amoscato, Andrew A et al. (2017) Gas Cluster Ion Beam Time-of-Flight Secondary Ion Mass Spectrometry High-Resolution Imaging of Cardiolipin Speciation in the Brain: Identification of Molecular Losses after Traumatic Injury. Anal Chem 89:4611-4619
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
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
Li, Xianchan; Mohammadi, Amir Saeid; Ewing, Andrew G (2016) Single cell amperometry reveals curcuminoids modulate the release of neurotransmitters during exocytosis from PC12 cells. J Electroanal Chem (Lausanne) 781:30-35
Tarolli, Jay Gage; Bloom, Anna; Winograd, Nicholas (2016) Multimodal image fusion with SIMS: Preprocessing with image registration. Biointerphases 11:02A311
Tian, Hua; Maci??ek, 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

Showing the most recent 10 out of 26 publications