Single-element Resolution Research Articles

From SuperTIGER to TIGERISS

The Trans-Iron Galactic Element Recorder (TIGER) family of instruments is optimized to measure the relative abundances of the rare, ultra-heavy galactic cosmic rays (UHGCRs) with atomic number (Z) Z ≥ 30. Observing the UHGCRs places a premium on exposure that the balloon-borne SuperTIGER achieved with a large area detector (5.6 m2) and two Antarctic flights totaling 87 days, while the smaller (∼1 m2) TIGER for the International Space Station (TIGERISS) aims to achieve this with a longer observation time from one to several years. SuperTIGER uses a combination of scintillator and Cherenkov detectors to determine charge and energy. TIGERISS will use silicon strip detectors (SSDs) instead of scintillators, with improved charge resolution, signal linearity, and dynamic range. Extended single-element resolution UHGCR measurements through 82Pb will cover elements produced in s-process and r-process neutron capture nucleosynthesis, adding to the multi-messenger effort to determine the relative contributions of supernovae (SNe) and Neutron Star Merger (NSM) events to the r-process nucleosynthesis product content of the galaxy.

Co-culture hydrogel micro-chamber array-based plate for anti-tumor drug development at single-element resolution

In response to the need for reliable cellular models that reflect complex tumor microenvironmental properties, and enable more precise testing of anti-cancer therapeutics effects on humans, a co-culture platform for in-vitro model that enhances the physiology of breast cancer (BC) microenvironment is presented. A six well imaging plate wherein each macro-well contains several separate compartments was designed. Three-dimensional (3D) cancer spheroids are generated and cultured in the inner compartment which is embossed with an array of nano-liter micro-chambers made of hydrogel. Stromal cells are cultured in the outer chambers. The two cell types are cultured side-by-side, sharing a common space, thus enabling extra-cellular communication via secreted molecules. As proof of concept, a model of BC tumor microenvironment was recapitulated by co-cultivating 3D MCF7 spheroids in the presence of tumor-associated macrophages (TAMs). The presence of TAMs induced an aggressive phenotype by promoting spheroid growth, enhancing survivin expression levels and enabling invasive behavior. Moreover, TAMs influenced the response of BC spheroids to cytotoxic treatment as well as hormonal drug therapy, and enhanced the effects of nitric oxide donor. The platform enables time-lapse imaging and treatment without losing spatial location of the measured spheroids, thereby allowing measurements and analysis at individual-object resolution in an easy and efficient manner.

Analysis of Cancer Cell Invasion and Anti-metastatic Drug Screening Using Hydrogel Micro-chamber Array (HMCA)-based Plates.

Cancer metastasis is known to cause 90% of cancer lethality. Metastasis is a multistage process which initiates with the penetration/invasion of tumor cells into neighboring tissue. Thus, invasion is a crucial step in metastasis, making the invasion process research and development of anti-metastatic drugs, highly significant. To address this demand, there is a need to develop 3D in vitro models which imitate the architecture of solid tumors and their microenvironment most closely to in vivo state on one hand, but at the same time be reproducible, robust and suitable for high yield and high content measurements. Currently, most invasion assays lean on sophisticated microfluidic technologies which are adequate for research but not for high volume drug screening. Other assays using plate-based devices with isolated individual spheroids in each well are material consuming and have low sample size per condition. The goal of the current protocol is to provide a simple and reproducible biomimetic 3D cell-based system for the analysis of invasion capacity in large populations of tumor spheroids. We developed a 3D model for invasion assay based on HMCA imaging plate for the research of tumor invasion and anti-metastatic drug discovery. This device enables the production of numerous uniform spheroids per well (high sample size per condition) surrounded by ECM components, while continuously and simultaneously observing and measuring the spheroids at single-element resolution for medium throughput screening of anti-metastatic drugs. This platform is presented here by the production of HeLa and MCF7 spheroids for exemplifying single cell and collective invasion. We compare the influence of the ECM component hyaluronic acid (HA) on the invasive capacity of collagen surrounding HeLa spheroids. Finally, we introduce Fisetin (invasion inhibitor) to HeLa spheroids and nitric oxide (NO) (invasion activator) to MCF7 spheroids. The results are analyzed by in-house software which enables semi-automatic, simple and fast analysis which facilitates multi-parameter examination.

GALACTIC COSMIC RAY ORIGINS AND OB ASSOCIATIONS: EVIDENCE FROM SuperTIGER OBSERVATIONS OF ELEMENTS 26Fe THROUGH 40Zr

ABSTRACT We report abundances of elements from 26Fe to 40Zr in the cosmic radiation measured by the SuperTIGER (Trans-Iron Galactic Element Recorder) instrument during 55 days of exposure on a long-duration balloon flight over Antarctica. These observations resolve elemental abundances in this charge range with single-element resolution and good statistics. These results support a model of cosmic ray origin in which the source material consists of a mixture of % material from massive stars and ∼81% normal interstellar medium material with solar system abundances. The results also show a preferential acceleration of refractory elements (found in interstellar dust grains) by a factor of ∼4 over volatile elements (found in interstellar gas) ordered by atomic mass (A). Both the refractory and volatile elements show a mass-dependent enhancement with similar slopes.

Hydrogel microstructure live-cell array for multiplexed analyses of cancer stem cells, tumor heterogeneity and differential drug response at single-element resolution.

Specific phenotypic subpopulations of cancer stem cells (CSCs) are responsible for tumor development, production of heterogeneous differentiated tumor mass, metastasis, and resistance to therapies. The development of therapeutic approaches based on targeting rare CSCs has been limited partially due to the lack of appropriate experimental models and measurement approaches. The current study presents new tools and methodologies based on a hydrogel microstructure array (HMA) for identification and multiplex analyses of CSCs. Low-melt agarose integrated with type I collagen, a major component of the extracellular matrix (ECM), was used to form a solid hydrogel array with natural non-adhesive characteristics and high optical quality. The array contained thousands of individual pyramidal shaped, nanoliter-volume micro-chambers (MCs), allowing concomitant generation and measurement of large populations of free-floating CSC spheroids from single cells, each in an individual micro-chamber (MC). The optical live cell platform, based on an imaging plate patterned with HMA, was validated using CSC-enriched prostate and colon cancer cell lines. The HMA methodology and quantitative image analysis at single-element resolution clearly demonstrates several levels of tumor cell heterogeneity, including morphological and phenotypic variability, differences in proliferation capacity and in drug response. Moreover, the system facilitates real-time examination of single stem cell (SC) fate, as well as drug-induced alteration in expression of stemness markers. The technology may be applicable in personalized cancer treatment, including multiplex ex vivo analysis of heterogeneous patient-derived tumor specimens, precise detection and characterization of potentially dangerous cell phenotypes, and for representative evaluation of drug sensitivity of CSCs and other types of tumor cells.

Cosmic ray composition at high energies: The TRACER project

The TRACER instrument ( Transition Radiation Array for Cosmic Energetic Radiation) is designed to measure the individual energy spectra of cosmic ray nuclei in long duration balloon flights. The large geometric factor of TRACER (5 m 2 sr) permits statistically significant measurements at particle energies well beyond 10 14 eV. TRACER identifies individual cosmic ray nuclei with single element resolution, and measures their energy over a wide range, from about 0.5 to 10,000 GeV/amu. This is accomplished with a gas detector system of 1600 single wire proportional tubes and plastic fiber radiators combined with plastic scintillators and acrylic Cerenkov counters. A 2-week flight in Antarctica in December 2003 has led to a measurement of the nuclear species oxygen to iron up to about 3000 GeV/amu. As an example, we shall present the energy spectrum and relative abundance for neon and discuss the implication of this result in the context of current models of acceleration and propagation of galactic cosmic rays. The instrument has been refurbished and flown on a second long duration balloon flight in Summer 2006. For this flight, the dynamic range of TRACER has been extended to permit inclusion of the lighter elements B, C, and N in the measurement.