Endgroup determination of synthetic polymers by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

AbstractElectrospray ionization (ESI) was performed on a Fourier transform ion cyclotron resonance mass spectrometer for the endgroup and monomer mass determination of three poly(oxyalkylene)s in the mass range of 400–8000 Da. A combined use of the multiple charge states observed with ESI, leads to a threefold increase in accuracy of the endgroup and monomer determination. The improvement is attributed to the increased number of datapoints used for the regression procedure, yielding more accurate results. Endgroup masses are determined with a mass error better than 5 and 75 millimass units for the molecular weight range of 400–4200 and 6200–8000 Da, respectively. A mass error of better than 1 millimass unit was observed for all monomer mass determinations. With ESI, endgroup and monomer masses have been determined for poly(ethylene glycol) oligomers with a mass higher than 8000 Da. This is almost two times higher than observed with matrix-assisted laser desorption/ionization on the same instrument

Spatially Resolved Immunometabolism to Understand Infectious Disease Progression

Infectious diseases, including those of viral, bacterial, fungal, and parasitic origin are often characterized by focal inflammation occurring in one or more distinct tissues. Tissue-specific outcomes of infection are also evident in many infectious diseases, suggesting that the local microenvironment may instruct complex and diverse innate and adaptive cellular responses resulting in locally distinct molecular signatures. In turn, these molecular signatures may both drive and be responsive to local metabolic changes in immune as well as non-immune cells, ultimately shaping the outcome of infection. Given the spatial complexity of immune and inflammatory responses during infection, it is evident that understanding the spatial organization of transcripts, proteins, lipids, and metabolites is pivotal to delineating the underlying regulation of local immunity. Molecular imaging techniques like mass spectrometry imaging and spatially resolved, highly multiplexed immunohistochemistry and transcriptomics can define detailed metabolic signatures at the microenvironmental level. Moreover, a successful complementation of these two imaging techniques would allow multi-omics analyses of inflammatory microenvironments to facilitate understanding of disease pathogenesis and identify novel targets for therapeutic intervention. Here, we describe strategies for downstream data analysis of spatially resolved multi-omics data and, using leishmaniasis as an exemplar, describe how such analysis can be applied in a disease-specific context

A novel dual ionization modality source for infrared laser ablation post-ionization mass spectrometry imaging to study fungicide metabolism and transport

We present a novel probe design for ambient laser-based mass spectrometry imaging combining electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) in a single probe, compatible with a commercial laser ablation electrospray ionization (LAESI) instrument. Here we describe the probe design considerations and features, as well as an in-house developed data processing routine designed to extract accurate mass spectrometry imaging data from ambient laser ablation post-ionization experiments. We characterize the probe performance in both APCI and ESI mode on a selection of compounds and show improved pixel-to-pixel repeatability for LA-APCI as compared to LAESI. We apply the dual ionization probe in APCI mode in a time series experiment to monitor agrochemicals on tomato plants. We investigate the translocation of fungicide isotianil and one of its metabolites, anthranilonitrile, by mass spectrometry imaging over a period of two weeks after application on a leaf surface. LA-APCI-MSI shows translocation of anthranilonitrile from treated leaves towards non-treated leaves. In summary, we demonstrate that LA-APCI imaging is a valuable addition to the ambient mass spectrometry toolbox, with particular advantages for imaging experiments across a variety of compounds

ToF-SIMS parallel imaging MS/MS of lipid species in thin tissue sections

Unambiguous identification of detected species is essential in complex biomedical samples. To date, there are not many mass spectrometry imaging techniques that can provide both high spatial resolution and identification capabilities. A new and patented imaging tandem mass spectrometer, exploiting the unique characteristics of the nanoTOF II (Physical Electronics, USA) TOF-SIMS TRIFT instrument, was developed to address this. Tandem mass spectrometry is based on the selection of precursor ions from the full secondary ion spectrum (MS1), followed by energetic activation and fragmentation, and collection of the fragment ions to obtain a tandem MS spectrum (MS2). The PHI NanoTOF II mass spectrometer is equipped with a high-energy collision induced dissociation (CID) fragmentation cell as well as a second time-of-flight analyzer developed for simultaneous ToF-SIMS and tandem MS imaging experiments. We describe here the results of a ToF-SIMS imaging experiment on a thin tissue section of an infected zebrafish as a model organism for tuberculosis. The focus is on the obtained ion distribution plot of a fatty acid as well as its identification by tandem mass spectrometry

Mass spectrometry imaging reveals flavor distribution in edible mushrooms

The spatial distribution of molecules and compounds responsible for the flavor profile of edible button mushrooms (Agaricus bisporous) has never been determined. The food industry is interested in knowing the localization of these compounds. Such knowledge would enable extraction of flavor compounds from a particular regions of the mushroom, which is safer for consumption compared to alternatives such as synthetic flavoring agents. The present study utilizes matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI), to determine the spatial distribution of flavor compounds in a mushroom. As MALDI-MSI requires very thin sections, a sample preparation protocol was optimized and sectioning fresh frozen mushrooms at 35 µm thickness was considered the best method to evaluate the distribution of flavor compounds. Further, the effect of heat on the spatial distribution of flavor compounds was investigated by heating whole mushrooms to 140 ? prior to sectioning. Heating reduced the water content of the mushroom and thus enabled the generation of even-thinner 17 µm thick sections. MALDI-MSI measurements performed on underivatized and on-tissue derivatized fresh frozen and heat-treated mushroom sections elucidated the spatial distribution of several flavor-related compounds

Automated, Feature-Based Image Alignment for High-Resolution Imaging Mass Spectrometry of Large Biological Samples

High-resolution imaging mass spectrometry of large biological samples is the goal of several research groups. In mosaic imaging, the most common method, the large sample is divided into a mosaic of small areas that are then analyzed with high resolution. Here we present an automated alignment routine that uses principal component analysis to reduce the uncorrelated noise in the imaging datasets, which previously obstructed automated image alignment. An additional signal quality metric ensures that only those regions with sufficient signal quality are considered. We demonstrate that this algorithm provides superior alignment performance than manual stitching and can be used to automatically align large imaging mass spectrometry datasets comprising many individual mosaic tiles