Post-ionization Secondary Neutral Mass Spectrometry Research Articles

Implementation and Optimization of Large Gas Cluster Laser Post-Ionization Secondary Neutral Mass Spectrometry for Molecular Analysis

In this study, we describe for the first time a new combined technique for molecular surface analysis using large argon gas clusters as primary ions for laser postionization secondary neutral mass spectrometry (Laser-SNMS). This new technique was investigated on polymer polystyrene (PS) surfaces using pulsed Ar2000+ ions with 20 keV energy and a 157 nm laser beam for postionization. To optimize the ion yield, time-of-flight (ToF) distributions of characteristic sputtered neutral PS fragments were determined. The data could be fitted well with Maxwell–Boltzmann distributions. The data show that the maxima of the individual ToF distributions of the sputtered neutrals move to higher delay time values with increasing mass, which directly correlates to a decrease of velocity of the sputtered neutrals with increasing mass. The delay time dependence on mass could be fitted with a quadratic curve, thus indicating a constant mean kinetic energy for the sputtered molecules. The ion yields obtained with large gas cl...

Improved 3D-imaging of a sirolimus/probucol eluting stent coating using laser postionization secondary neutral mass spectrometry and time-of-flight secondary ion mass spectrometry

Implantable drug delivery systems that provide controlled and sustained release of a therapeutic agent are used in a wide variety of applications. Drug eluting stents, which are used to treat coronary artery disease, are among the most widespread of these devices, with an estimated 3 × 106 implants annually worldwide. Controlling the rate of drug release from these devices relies on precise control of the three dimensional (3D)-distribution of the drug, so methods for measuring this distribution are of great importance. The aims of this work were to determine how 3D-imaging of polymer-free sirolimus/probucol drug eluting stent coatings could be improved through the use of laser postionization secondary neutral mass spectrometry (Laser-SNMS) and Ar cluster sputtering with time-of-flight secondary ion mass spectrometry (ToF-SIMS) and to optimize conditions for this analysis. In this study, 3D-imaging of a sirolimus/probucol dual drug eluting stent has been investigated using Laser-SNMS and ToF-SIMS. Laser-SNMS studies of pure sirolimus and probucol were undertaken using 30 keV Bi3+ primary ions and a 157 nm excimer postionization laser. Under optimal conditions, a greater than 100-fold increase in detected ion yield was observed for Laser-SNMS when compared to ToF-SIMS, although ToF-SIMS provided equal or greater yields for higher mass characteristic ions. Although the optimal laser power density for detecting probucol (5 × 106 W/cm2) was significantly lower than the optimum for sirolimus (7 × 107 W/cm2), an intermediate laser power density of 1 × 107 W/cm2 was sufficient to allow imaging of both drugs. Using individual selected ion signals, ToF-SIMS and Laser-SNMS produced similar images of the two drug species. When using, however, a multivariate approach (maximum autocorrelation factors), Laser-SNMS provided significant improvements in image contrast and small area detection when compared to ToF-SIMS. Following optimization of the technique, 3D-images of the dual drug eluting stent coating were obtained using 10 keV Ar2000+ cluster ions for sputtering and 30 keV Bi3+ cluster ions for analysis for both ToF-SIMS and Laser-SNMS. This work demonstrates the advantages of Laser-SNMS for 3D-imaging of pharmaceutical devices, which has not been previously published. Both ToF-SIMS and Laser-SNMS revealed that the outermost surface of the drug eluting coating contained pure sirolimus to a depth of a few tens of nanometers, with a few channels of sirolimus extending to a depth of around 1 μm. Below about 1 μm, the two drugs were uniformly mixed. Using the 10 keV Ar2000+ sputter beam, the authors were able to sputter through the complete drug coating (∼6 μm) without observing any accumulated damage in the organic layer. The two techniques showed complementary strengths: ToF-SIMS offers faster data collection and better detected ion yield for larger characteristic ions than Laser-SNMS, and Laser-SNMS offers significantly enhanced detected ion yield for smaller fragment ions, allowing for improved image contrast and resolution of smaller features.

ToF-SIMS and Laser-SNMS Imaging of Heterogeneous Topographically Complex Polymer Systems.

Heterogeneous polymer coatings, such as those used in organic electronics and medical devices, are of increasing industrial importance. In order to advance the development of these types of systems, analytical techniques are required which are able to determine the elemental and molecular spatial distributions, on a nanometer scale, with very high detection efficiency and sensitivity. The goal of this study was to investigate the suitability of laser postionization secondary neutral mass spectrometry (Laser-SNMS) with a 157 nm postionization laser beam to image structured polymer mixtures and compare the results with time-of-flight secondary ion mass spectrometry (ToF-SIMS) measurements using Bi3+ primary ions. The results showed that Laser-SNMS is better suited than ToF-SIMS for unambiguous detection and submicrometer imaging of the wide range of polymers investigated. The data also showed that Laser-SNMS has the advantage of being much more sensitive (in general higher by more than an order of magnitude and peaking at up to 3 orders of magnitude) than ToF-SIMS while also showing superior performance on topographically complex structured insulating surfaces, due to significantly reduced field effects and a higher dynamic range as compared to ToF-SIMS. It is concluded that Laser-SNMS is a powerful complementary technique to ToF-SIMS for the analysis of heterogeneous polymers and other complex structured organic mixtures, providing submicrometer resolution and high sensitivity.

ToF-SIMS and laser-SNMS analysis of Madin-Darby canine kidney II cells with silver nanoparticles using an argon cluster ion beam.

The use of nanoparticles is one of the fastest expanding fields in industrial as well as in medical applications, owing to their remarkable characteristics. Silver nanoparticles (AgNPs) are among the most-commercialized nanoparticles because of their antibacterial effects. Laser postionization secondary neutral mass spectrometry (laser-SNMS) and time-of-flight secondary ion mass spectrometry in combination with argon cluster ion sputtering was used for the first time to investigate the effects of AgNPs on Madin-Darby canine kidney (MDCK) II cells. Depth profiles and high-resolution three dimensional (3D) images of nanoparticles and organic compounds from cells were obtained using an Ar cluster ion beam for sputtering and Bi3 (+) primary ions for the analysis. The 3D distribution of AgNPs and other organic compounds in MDCK II cells could be readily detected with very high efficiency, sensitivity, and submicron lateral resolution. The argon cluster ion beam is well suited for the sputtering of biological samples. It enables a high sample removal rate along with low molecular degradation. The outer membrane, the cytoplasm, and the nuclei of the cells could be clearly visualized using the signals PO(+) and C3H8N(+) or CN(+) and C3H8N(+). The laser-SNMS images showed unambiguously that AgNPs are incorporated by MDCK II cells and often form silver aggregates with a diameter of a few micrometers, mainly close to the outside of the cell nuclei.

Application of Laser Postionization Secondary Neutral Mass Spectrometry/Time-of-Flight Secondary Ion Mass Spectrometry in Nanotoxicology: Visualization of Nanosilver in Human Macrophages and Cellular Responses

Silver nanoparticles (SNP) are the subject of worldwide commercialization because of their antimicrobial effects. Yet only little data on their mode of action exist. Further, only few techniques allow for visualization and quantification of unlabeled nanoparticles inside cells. To study SNP of different sizes and coatings within human macrophages, we introduce a novel laser postionization secondary neutral mass spectrometry (Laser-SNMS) approach and prove this method superior to the widely applied confocal Raman and transmission electron microscopy. With time-of-flight secondary ion mass spectrometry (TOF-SIMS) we further demonstrate characteristic fingerprints in the lipid pattern of the cellular membrane indicative of oxidative stress and membrane fluidity changes. Increases of protein carbonyl and heme oxygenase-1 levels in treated cells confirm the presence of oxidative stress biochemically. Intriguingly, affected phagocytosis reveals as highly sensitive end point of SNP-mediated adversity in macrophages. The cellular responses monitored are hierarchically linked, but follow individual kinetics and are partially reversible.

Laser secondary neutral mass spectrometry for copper detection in micro‐scale biopsies

Disease progression and clinical diagnostics of a number of hereditable metabolic diseases are determined by organ involvement in disturbed deposition of certain molecules. Current clinical imaging is unable to visualize this maldistribution with sufficient specificity and sensitivity, such as in Wilson's disease. The quest for understanding cellular Cu distribution in these patients requires element- and molecule-specific images with nanometer-scale spatial resolution. We have used a new cryo-mass spectrometric instrument with an integrated cryosectioning chamber for preparation and analysis of frozen hydrated samples of Wilson's disease tissue. With laser post-ionization secondary neutral mass spectrometry (laser-SNMS), we were able to image Cu and other intrinsic elements and molecules in less than 1 mg of frozen hydrated liver tissue from a murine model of Wilson's disease. A 40-50 times higher Cu concentration was measured in the disease tissue as compared to the control mouse. Furthermore, major histomorphological changes were observed using this advanced nano-science tool. The results showed that the combination of in-vacuum cryosectioning and cryo-laser-SNMS technologies is particularly well suited for identifying specific cell structures and imaging trace element concentrations with subcellular resolution and upper-parts-per-billion sensitivity in biological samples. This technology can provide a novel diagnostic tool for clinical applications in various diseases involving trace elements.

Laser postionization secondary neutral mass spectrometry in tissue: a powerful tool for elemental and molecular imaging in the development of targeted drugs

The exact intracellular localization and distribution of molecules and elements becomes increasingly important for the development of targeted therapies and contrast agents. We show that laser postionization secondary neutral mass spectrometry (laser-SNMS) is well suited to localize particular elements and small molecules with subcellular spatial resolution applying the technique exemplary to Boron Neutron Capture Therapy (BNCT). We showed in a murine sarcoma that the drugs used for clinical BNCT, namely l-para-boronophenylalanine (700 mg/kg body weight i.p.) and sodium mercaptoundecahydro-closo-dodecaborate (200 mg/kg body weight i.p.), transport the therapeutic agent (10)B into the cytoplasm and into the nucleus itself, the most sensitive area of the cell. Sodium mercaptoundecahydro-closo-dodecaborate distributes (10)B homogeneously and l-para-boronophenylalanine heterogeneously. When combining laser-SNMS with prompt gamma-ray analysis as a screening technique, strategies for BNCT can be elaborated to develop new drugs or to improve the use of existing drugs on scientifically based evidence. The study shows the power of laser-SNMS in the early stages of drug development, also outside BNCT.

Possibilities and limitations of high-resolution mass spectrometry in life sciences

We have applied time-of-flight secondary ion mass spectrometry (ToF-SIMS) and laser post-ionization secondary neutral mass spectrometry (laser-SNMS) to examine the immobilization process of PNA and its hybridization capability to unlabeled complementary DNA fragments, to characterize immobilized proteins, and to image intrinsic elements and molecules with subcellular spatial resolution in different types of frozen non-dehydrated biological samples. The possibilities and limitations of ToF-SIMS and laser-SNMS for imaging elements and molecules in biological samples are discussed. Furthermore possibilities for enhancing the detection sensitivity by using polyatomic and cluster primary ions and different laser post-ionization schemes, as well as ways of obtaining 3D molecular images from biological samples are described. The data shows that both ToF-SIMS and laser-SNMS are capable of imaging elements and molecules in complex biological samples and that they are very valuable tools in advancing applications in life sciences. It was found that cluster-ion bombardment is very useful for enhancing the molecular yield, while laser-SNMS resulted in much higher detection sensitivity for elements and specific molecules and is particularly well suited for imaging ultra-trace element concentrations in biological samples.

Mass spectrometry on the nanoscale with ion sputtering based techniques: What is feasible

The potential of ion sputtering based mass spectrometry applied to materials characterization on the nanometer scale is discussed. Analytical approaches and required instrumental capabilities are outlined, and the current state-of-the-art is summarized. A new generation of analytical instruments specifically optimized for laser post-ionization secondary neutral mass spectrometry has been developed at Argonne National Laboratory (ANL). Experimentally verified (or anticipated after near-future upgrades) analytical capabilities of these instruments, capable of quantitative analysis at the nanometer-scale, are reported and compared to secondary ion mass spectrometry.

Mass spectrometric characterization of elements and molecules in cell cultures and tissues

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) and laser post-ionization secondary neutral mass spectrometry (laser-SNMS) have been used to image and quantify targeted compounds, intrinsic elements and molecules with subcellular resolution in single cells of both cell cultures and tissues. Special preparation procedures for analyzing cell cultures and tissue materials were developed. Cancer cells type MeWo, incubated with boronated compounds, were sandwiched between two substrates, cryofixed, freeze-fractured and freeze-dried. Also, after injection with boronated compounds, different types of mouse tissues were extracted, prepared on a special specimen carrier and plunged with high velocity into LN 2-cooled propane for cryofixation. After trimming, these tissue blocks were freeze-dried. The measurements of the K/Na ratio demonstrated that for both cell cultures and tissue materials the special preparation techniques used were appropriate for preserving the chemical and structural integrity of the living cell. The boron images show inter- and intracellular boron signals with different intensities. Molecular images show distinct features partly correlated with the cell structure. A comparison between laser-SNMS and ToF-SIMS showed that especially laser-SNMS is particularly well-suited for identifying specific cell structures and imaging ultratrace element concentrations in tissues.

Laser post-ionization secondary neutral mass spectrometry for ultra-trace analysis of samples from space return missions

A new generation of secondary neutral mass spectrometry (SNMS) instruments has been developed that is especially designed for laser post-ionization (LPI). These instruments combine high useful yield and high background discrimination. Results presented here demonstrate that these instruments can detect one in every four atoms removed from a samples surface – a greater than one order of magnitude improvement over current large frame secondary ion mass spectrometry instruments. Because of their high sensitivity, these new LPI-SNMS instruments are especially amenable to analysis of samples of limited size and rare one-of-a-kind samples. Such an application is analysis of samples returned to Earth from space by the Genesis and Stardust Discovery missions of NASA.

A new time-of-flight instrument for quantitative surface analysis

A new generation of time-of-flight mass spectrometers that implement ion sputtering and laser desorption for probing solid samples and can operate in regimes of laser post-ionization secondary neutral mass spectrometry and secondary ion mass spectrometry is being developed at Argonne National Laboratory. These new instruments feature novel ion optical systems for efficient extraction of ions from large laser post-ionization volumes and for lossless transport of these ions to detectors. Another feature of this design is a new in-vacuum all-reflecting optical microscope with 0.5-μm resolution. Advanced ion and light optics and three ion sources, including a liquid metal ion gun (focusable to 50 nm) and a low energy ion gun, give rise to an instrument capable of quantitative analyses of samples for the most challenging applications, such as determining elemental concentrations in shallow implants at ultra-trace levels (for example, solar wind samples delivered by NASA Genesis mission) and analyzing individual sub-micrometer particles on a sample stage (such as, interstellar dust delivered by NASA Stardust mission). Construction of a prototype instrument has been completed and testing is underway. A more advanced instrument of similar design is under construction. The overall design of the new instrument and the innovations that make it unique are outlined. Results of the first tests to characterize its analytical capabilities are presented also.

Relative elemental sensitivity factors in non-resonant laser-SNMS.

Using a reflectron time-of-flight mass spectrometer, the ionization process in non-resonant Laser postionization Secondary Neutral Mass Spectrometry (SNMS) has been investigated. In particular, the postionization efficiencies (PIE) achieved by multi photon and single photon absorption have been compared by ionizing ten elements sputtered from a NIST standard reference material by excimer laser radiation of 248 nm, 193 nm and 157 nm. Only in the case of single photon ionization (SPI) the measured laser intensity dependence of the PIE can be understood quantitatively in terms of corresponding theory. From the results, absolute values of the SPI cross sections have been evaluated for atoms of nine elements, which show a total variation over about two orders of magnitude. Furthermore, even in the regime of high laser intensity, where the ionization of all atoms is completely saturated, different elements have been detected with relative sensitivity factors which scatter over about one order of magnitude. This has been attributed to element dependent variations of the effective ionization volume which are caused by the different kinetic energy and angular distributions of different sputtered atoms.