Combined Ion Beam Irradiation Platform and 3D Fluorescence Microscope for Cellular Cancer Research

To improve particle radiotherapy, we need a better understanding of the biology of radiation effects, particularly in heavy ion radiation therapy, where global responses are observed despite energy deposition in only a subset of cells. Here, we integrated a high-speed swept confocally-aligned planar excitation (SCAPE) microscope into a focused ion beam irradiation platform to allow real-time 3D structural and functional imaging of living biological samples during and after irradiation. We demonstrate dynamic imaging of the acute effects of irradiation on 3D cultures of U87 human glioblastoma cells, revealing characteristic changes in cellular movement and intracellular calcium signaling following ionizing irradiation

Sputtering of Bi and Preferential Sputtering of an Inhomogeneous Alloy

Angular distributions and total yields of atoms sputtered from bismuth targets by normally incident 10 keV -50 keV Ne+ and Ar+ ions have been measured both experimentally and by computer simulation. Polycrystalline Bi targets were used for experimental measurements. The sputtered atoms were collected on high purity aluminum foils under ultra-high vacuum conditions, and were subsequently analyzed using Rutherford backscattering spectroscopy. The Monte-Carlo based SRIM code was employed to simulate angular distributions of sputtered Bi atoms and total sputtering yields of Bi to compare with experiment. The measured sputtering yields were found to increase with increasing projectile energy for normally incident 10 keV - 50 keV Ne+ and Ar+ ions. The shapes of the angular distributions of sputtered Bi atoms demonstrated good agreement between experiment and simulation in the present study. The measured and simulated angular distributions of sputtered Bi exhibited an over-cosine tendency. The measured value of the degree of this over-cosine nature was observed to increase with increasing incident Ne+ ion energy, but was not strongly dependent on incident Ar+ ion energy. The differential angular sputtering yield and partial sputtering yields due to Ar ion bombardment of an inhomogeneous liquid Bi:Ga alloy have been investigated, both experimentally and by computer simulation. Normally incident 25 keV and 50 keV beams of Ar+ were used to sputter a target of 99.8 at% Ga and 0.2 at% Bi held at 40° C in ultra-high vacuum (UHV), under which conditions the alloy is known to exhibit extreme Gibbsian surface segregation that produces essentially a monolayer of Bi atop the bulk liquid. Angular distributions of sputtered neutrals and partial sputtering yields obtained from the conversion of areal densities of Bi and Ga atoms on collector foils were determined. The Monte-Carlo based SRIM code was employed to simulate the experiment and obtain the angular distribution of sputtered components. The angular distribution of sputtered Ga atoms, originating from underneath the surface monolayer, was measured to be sharply peaked in angle about the surface normal direction compared to the Bi atoms originating from surface monolayer. The simulation study produced contradicting results, where the species originating from surface monolayer was strongly peaked around the surface normal compared to the species originating from beneath the surface monolayer

Sequestration of Ca in simian nasal mucosa : Determination of Ca molarity in ex vivo tissue by simultaneous off-axis Scanning Transmission Ion Microscopy, Particle Induced X-Ray Emission and Elastic Backscattering Spectrometry

A technique to determine the molar concentration of elements in ex vivo tissue sections by Particle Induced X-ray Emission (PIXE) has been developed. The method is based on simultaneous off-axis scanning transmission ion microscopy (OA-STIM) and Elastic Backscattering Spectrometry (EBS) measurement of the sample thickness and major element (H, C, N, O) composition. The method was applied to determine the molarity of localised Ca concentration hot-spots in the outer epithelium tissue of nasal mucosa of a rhesus macaque subject infected with simian immunodeficiency virus (SIV). The results show Ca sequestration in concentration hot-spots and outer epithelial tissue that significantly exceeded the Ca concentration in the surrounding tissues. This may originate from mineralisation and/or Ca enhancement in goblet cells

Machine learning approach for quantitative biodosimetry of partial-body or total-body radiation exposures by combining radiation-responsive biomarkers

Abstract During a large-scale radiological event such as an improvised nuclear device detonation, many survivors will be shielded from radiation by environmental objects, and experience only partial-body irradiation (PBI), which has different consequences, compared with total-body irradiation (TBI). In this study, we tested the hypothesis that applying machine learning to a combination of radiation-responsive biomarkers (ACTN1, DDB2, FDXR) and B and T cell counts will quantify and distinguish between PBI and TBI exposures. Adult C57BL/6 mice of both sexes were exposed to 0, 2.0–2.5 or 5.0 Gy of half-body PBI or TBI. The random forest (RF) algorithm trained on ½ of the data reconstructed the radiation dose on the remaining testing portion of the data with mean absolute error of 0.749 Gy and reconstructed the product of dose and exposure status (defined as 1.0 × Dose for TBI and 0.5 × Dose for PBI) with MAE of 0.472 Gy. Among irradiated samples, PBI could be distinguished from TBI: ROC curve AUC = 0.944 (95% CI: 0.844–1.0). Mouse sex did not significantly affect dose reconstruction. These results support the hypothesis that combinations of protein biomarkers and blood cell counts can complement existing methods for biodosimetry of PBI and TBI exposures

Biomarker integration for improved biodosimetry of mixed neutron + photon exposures

Abstract There is a persistent risk of a large-scale malicious or accidental exposure to ionizing radiation that may affect a large number of people. Exposure will consist of both a photon and neutron component, which will vary in magnitude between individuals and is likely to have profound impacts on radiation-induced diseases. To mitigate these potential disasters, there exists a need for novel biodosimetry approaches that can estimate the radiation dose absorbed by each person based on biofluid samples, and predict delayed effects. Integration of several radiation-responsive biomarker types (transcripts, metabolites, blood cell counts) by machine learning (ML) can improve biodosimetry. Here we integrated data from mice exposed to various neutron + photon mixtures, total 3 Gy dose, using multiple ML algorithms to select the strongest biomarker combinations and reconstruct radiation exposure magnitude and composition. We obtained promising results, such as receiver operating characteristic curve area of 0.904 (95% CI: 0.821, 0.969) for classifying samples exposed to ≥ 10% neutrons vs. < 10% neutrons, and R2 of 0.964 for reconstructing photon-equivalent dose (weighted by neutron relative biological effectiveness) for neutron + photon mixtures. These findings demonstrate the potential of combining various -omic biomarkers for novel biodosimetry

Dynamic Analysis of major elements in biological tissue validating quantification of trace life elements in MeV ion beam microscopy

MeV ion microprobe measurements of the lesser and trace life element concentrations in biological tissues are challenging because of complex spatial inhomogeneities in these types of samples. Measurements on ex vivo tissue sections require determination of the matrix element composition and the tissue section thickness. For these reasons, in this work, we adapted the Dynamic Analysis approach known from literature, to interpret the MeV ion microscopy data to determine concentrations of H, C, O, and N as well as the thickness of different tissue regions in Non Human Primate (NHP) mesenteric lymph node section. The results showed no strong variations of the matrix element contents regardless of section thickness variations in the tissue. The matrix information was used to quantify total-Ca molarities and a significant ∼30 mM Ca concentration hotspot was observed at the edge of sinus structure in the mesenteric lymph node as compared to the 3-4 mM total-Ca levels in the surrounding tissues. Thus, MeV ion microprobe imaging combined with dynamic analysis comprise a novel chemometric approach paving a way for quantitative analysis of similarly complicated animal and plant biological tissue sections