In this work the chemical analysis of biological cells and tissues with time-of-flight secondary ion mass spectrometry (ToF-SIMS) was explored. ToF-SIMS has the ability to obtain a mass spectrum with submicron spatial resolution for imaging and is extremely surface sensitive. ToF-SIMS for biological sample analysis is still an emerging field, so the development and characterization of novel sample preparation and analysis methods is key to acquiring useable information. In this work, three different methods to prepare NIH/3T3 fibroblasts were investigated: chemically fixed, freeze-dried and frozen-hydrated. Chemical fixation followed by rinsing removed a majority of intracellular Cl-, improving the secondary ion yields of all organic positively charged secondary ions an average of 2.6x. Damage cross sections were reduced during frozen-hydrated analysis, improving the secondary ion yields of higher mass organic fragments. In a separate experiment, accurate 3D reconstructions of NIH/3T3 fibroblasts were produced. A simple z-correction was applied to the data cube, and the biggest assumption for that correction was validated. An intracellular lipid-rich region surrounding the nucleus was visualized. ToF-SIMS applied to two different breast cancer systems. In the first, eight human breast cancer cell lines were distinguished from one another using mass spectra and principal component analysis (PCA). Not only was PCA to distinguish the cell lines form one another, it also highlighted the largest sources of variance between the cells. Phosphocholine, fatty acids, cholesterol and diacylglycerols (DAGs) were identified as key peaks. The identification of these species indicate that differences in lipid metabolism play an important role in separating the cell types from one another. Breast cancer tumor tissues were also investigated. Data from four tumors was collected. PCA applied to the spectra distinguished the four tissues from one another. Imaging PCA determined the largest sources of variance within an analysis area. Structures were identified by PCA that matched structures observed in serial-sectioned, conventionally-stained slices, and other domains that were not visible in the conventionally-stained slices. As with the breast cancer cell lines, phosphocholine, fatty acids, DAGs, cholesterol and vitamin were found to be large sources of variance, indicating lipid metabolism plays in important role in tumor differentiation.

This second edition details new and updated chapters on key methodologies and breakthroughs in the mass spectrometry imaging (MSI) field. Chapters guide readers through nano-Desorption Electrospray Ionisation (nDESI), Matrix Assisted Laser Desorption Ionisation-2 (MALDI-2), Laser Ablation - Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) ,Imaging Mass Cytometry (IMC) with a variety of diverse samples including eye tissue, crop analysis, 3D cell culture models, and counterfeit goods analysis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Imaging Mass Spectrometry: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

This thesis project consisted of two separate projects which were connected by roots in biological analysis. The first was to characterize the release kinetics of a protein from a biodegradable polymer membrane which had not previously been formulated on a scale large enough for clinical medicine. This scaled up membrane was assessed using Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS) and fluorescence spectroscopy for physical properties such as composition and release kinetics but was also analyzed using optical microscopy methods for biocompatibility. The second was to use of ToF-SIMS chemical imaging to determine the distribution of a drug called methylselenocysteine (MSC) in xenograft tumor tissue. MSC had previously been found to increase tumor vasculature which will in turn increase the distribution of CPT-11 throughout the tumor. This type of study had not previously been attempted with the ToF-SIMS chemical imaging method.^Based on the work done on both of these projects, three chapters are included in this thesis document and are summarized below. A new, low mass aggregation pattern was observed using ToF-SIMS for solid Aerosol-OT (AOT) thin films cast from concentrations above the critical micelle concentration (CMC) (10-3 - 10-5 M in chloroform). Conventional methods typically used to calculate the aggregation number of AOT include optical methods such as visible spectroscopy, dynamic light scattering, or infrared spectroscopy which use the appearance (or disappearance) of a pattern to discern the CMC. These methods all use a secondary molecule such as water to determine when a physical change in the AOT has occurred.^ToF-SIMS analysis allows for the direct analysis of an ion fragmentation pattern specific to the solution concentration of AOT to determine the CMC. Two molecularly distinct polymer membranes based on optimization of KGF activity were formulated to release biologically active KGF into solution. The protein that was used for this study is Keratinocyte Growth Factor (KGF). The membrane was formulated with an anionic surfactant AOT, which was hypothesized to contain the protein in an aqueous environment thereby preserving the activity during the formulation and delivery process. Each of these membranes was found to release a biologically relevant concentration of KGF and was able to sustain cell growth/adherence in vitro. ToF-SIMS chemical imaging was conducted using tissue that had been treated first with methylselenocystine (MSC) followed by the anticancer drug CPT-11.^ToF-SIMS analysis found that tumor and liver tissue that had been treated with MSC had characteristic ion signals that could be associated with this drug. Imaging with ToF-SIMS was confirmed with fluorescence microscopy and found that the drug segregated to the vasculature of the liver and tumor tissue.

Applications of Mass Spectrometry Imaging to Cancer, the latest volume in the Advances in Cancer Research provides invaluable information on the exciting and fast-moving field of cancer research. This volume presents original reviews on applications of mass spectrometry imaging to cancer. - Provides information on cancer research - Offers outstanding and original reviews on a range of cancer research topics - Serves as an indispensable reference for researchers and students alike

The resurgence of time-of-flight mass spectrometry (TOF-MS) has had its origin in the simplicity of construction and application of such instruments together with the high transmission and the great increase in resolution that has been achieved. The instrument lends itself naturally to a coupling with pulsed laser sources, though this is not a prerequisite. It also affords a time resolution far beyond that traditionally achieved with mass spectrometric rapid scan techniques - a recent example being the real-time analysis of a multi-component mixture from an automobile exhaust. Furthermore, the mass range appears to be extremely large: mass up to 500 kDa and beyond what is being readily measured in the laboratory today. The present set of contributions attempts to give a survey of current applications from many of the active groups in the field. A variety of new applications are considered which are no doubt just the beginning of large new areas of application. By presenting this work in book form it is hoped that it will be of help to the many groups intending to initiate work in this rapidly expanding new area of mass spectrometry.

Due to its enormous sensitivity and ease of use, mass spectrometry has grown into the analytical tool of choice in most industries and areas of research. This unique reference provides an extensive library of methods used in mass spectrometry, covering applications of mass spectrometry in fields as diverse as drug discovery, environmental science, forensic science, clinical analysis, polymers, oil composition, doping, cellular research, semiconductor, ceramics, metals and alloys, and homeland security. The book provides the reader with a protocol for the technique described (including sampling methods) and explains why to use a particular method and not others. Essential for MS specialists working in industrial, environmental, and clinical fields.