Neutron microscopy. The low-damage imaging of specialized organic materials

It is shown that, insofar as radiation damage is concerned, transmission neutron microscopy using neutrons in the energy range approximately 0.0001-1.0 eV is extremely attractive for the imaging of specialized organic materials. By "specialized organic materials" is meant organic specimens composed entirely of specific isotopes that have been selected on the basis of their favorable properties with regard to radiation damage. In connection with such specimens, it is demonstrated that at a resolution of, for example, 100 A, neutrons will have an advantage over soft X-rays in terms of radiation damage, provided that the inherent (neutron) bright field image contrast turns out to be greater than 10(-5). Suggestions relating to (a) the comprehensive calculation of the radiation damage sustained by specialized organic specimens under slow neutron irradiation, (b) the construction of a theory of image formation in the neutron microscope, (c) the development of neutron lenses/focusing devices, and (d) the development of a brighter neutron source (essential for neutron microscopy) are outlined in some detail. The paper concludes with two appendices, which provide important background material.     

High-Contrast Observation of Unstained Proteins and Viruses by Scanning Electron Microscopy

Scanning electron microscopy (SEM) is an important tool for the nanometre-scale analysis of the various samples. Imaging of biological specimens can be difficult for two reasons: (1) Samples must often be left unstained to observe detail of the biological structures; however, lack of staining significantly decreases image contrast. (2) Samples are prone to serious radiation damage from electron beam. Herein we report a novel method for sample preparation involving placement on a new metal-coated insulator film. This method enables obtaining high-contrast images from unstained proteins and viruses by scanning electron microscopy with minimal electron radiation damage. These images are similar to those obtained by transmission electron microscopy. In addition, the method can be easily used to observe specimens of proteins, viruses and other organic samples by using SEM.     

Intense fusion neutron sources

The review describes physical principles underlying efficient production of free neutrons, up-to-date possibilities and prospects of creating fission and fusion neutron sources with intensities of 10¹⁵–10²¹ neutrons/s, and schemes of production and application of neutrons in fusion-fission hybrid systems. The physical processes and parameters of high-temperature plasmas are considered at which optimal conditions for producing the largest number of fusion neutrons in systems with magnetic and inertial plasma confinement are achieved. The proposed plasma methods for neutron production are compared with other methods based on fusion reactions in nonplasma media, fission reactions, spallation, and muon catalysis. At present, intense neutron fluxes are mainly used in nanotechnology, biotechnology, material science, and military and fundamental research. In the near future (10–20 years), it will be possible to apply high-power neutron sources in fusion-fission hybrid systems for producing hydrogen, electric power, and technological heat, as well as for manufacturing synthetic nuclear fuel and closing the nuclear fuel cycle. Neutron sources with intensities approaching 10²⁰ neutrons/s may radically change the structure of power industry and considerably influence the fundamental and applied science and innovation technologies. Along with utilizing the energy produced in fusion reactions, the achievement of such high neutron intensities may stimulate wide application of subcritical fast nuclear reactors controlled by neutron sources. Superpower neutron sources will allow one to solve many problems of neutron diagnostics, monitor nano-and biological objects, and carry out radiation testing and modification of volumetric properties of materials at the industrial level. Such sources will considerably (up to 100 times) improve the accuracy of neutron physics experiments and will provide a better understanding of the structure of matter, including that of the neutron itself.     

Comet assay to sense neutron 'fingerprint'

The suitability of comet assay to identify DNA damage induced by neutrons of varying energy was tested. For this purpose, monoenergetic neutrons from Hiroshima University Radiobiological Research Accelerator (HIRRAC) were used to induce DNA damage in irradiated human peripheral blood lymphocytes. The level of damage was computed as tail moment for different doses (0.125-1 Gy) and compared with the effects resulting from irradiation with (60)Co gamma. The neutron-irradiated cells exhibited longer comet tails consisting of tiny pieces of broken DNA in contrast to the streaking tails generated by (60)Co gamma. The peak biological effectiveness occurred at 0.37 and 0.57 MeV; a further increase or decrease in neutron energy led to a reduced RBE value. The RBE values, as measured by the comet assay, were 6.3, 5.4, 4.7, 4.3, 2.6, and 1.7 for 0.37, 0.57, 0.79, 0.186, 1, and 2.3 MeV neutrons. The lower RBE value obtained by the comet assay when compared to that for other biological end points is discussed. This study reports the usefulness of the alkaline comet assay for identifying DNA damage induced by neutrons of the same radiation weighting factor. The comet assay is a potential tool for use in neutron therapy, as well as a method for the rapid screening of samples from individuals accidentally exposed to radiation.     

Effect of Irradiation on Microorganisms in a Nuclear Reactor

The effects of irradiation in the JRR-1 (Japan Research Reactor No. 1, a homogeneous light water nuclear reactor; max. power, 50 KW) on microorganisms such as bacterial and fungal spores and yeast cells were investigated in comparison with those of ⁶⁰Co gamma radiation. As far as the lethal effect was concerned the dose rate of radiation in the experimental hole No. 16 of the JRR-1 was equivalent to 3.0×l0⁶~3.4×l0⁶ r/hr with ⁶⁰Co gamma radiation, and a ratio of the neutron effect to the gamma radiation effect on microorganisms in this hole was estimated to be approximately 3~5.4. The results different from those with gamma radiation were obtained in experiments such as post-NaCl treatment and spore germination. The considerable contribution of fast neutrons to the total biological effect of neutrons, in comparison with the thermal neutron effect, could be presumed from the microbiological experiments with the help of physical and chemical data. Morphological changes in post-irradiation growth were observed by means of phase contrast microscopy. No specific aftereffect was found.     

Alterations in dose and lineal energy spectra under different shieldings in the Los Alamos high-energy neutron field

Nuclear interactions of space radiation with shielding materials result in alterations in dose and lineal energy spectra that depend on the specific elemental composition, density and thickness of the material. The shielding characteristics of materials have been studied using charged-particle beams and radiation transport models by examining the risk reduction using the conventional dose-equivalent approach. Secondary neutrons contribute a significant fraction of the total radiation exposure in space. An experiment to study the changes in dose and lineal energy spectra by shielding materials was carried out at the Los Alamos Nuclear Science Center neutron facility. In the energy range of about 2 to 200 MeV, this neutron spectrum is similar in shape within a factor of about 2 to the spectrum expected in the International Space Station habitable modules. It is shown that with a shielding thickness of about 5 g cm(-2), the conventional radiation risk increases, in some cases by as much as a factor of 2, but decreases with thicknesses of about of 20 g cm(-2). This suggests that care must be taken in evaluating the shielding effectiveness of a given material by including both the charged-particle and neutron components of space radiation.     

Depletion of neural precursor cells after local brain irradiation is due to radiation dose to the parenchyma, not the vasculature

The underlying mechanisms associated with radiation-induced cognitive impairments remain elusive but may involve changes in hippocampal neural precursor cells. Proliferating neural precursor cells have been shown to be extremely sensitive to X rays, either from damage to the cells themselves and/or through microenvironmental factors, including the anatomical relationship with the microvasculature, which is altered by radiation. The neutron capture reaction in boron was used to determine whether the sensitivity of neural precursor cells was dominated by direct radiation effects or was mediated through changes in the microvasculature. Young adult rats were irradiated with X rays, neutrons only, or neutrons plus either mercapto-undecahydro-dodecaborane (BSH) or p-dihydroxyboryl-phenylalanine (BPA). BSH remains inside cerebral vessels, thereby limiting the neutron capture intravascularly; BPA readily passes into the parenchyma. One month after irradiation, cell proliferation and numbers of immature neurons were determined using immunohistochemistry. Results showed that (1) neural precursor cells and their progeny were decreased in a dose-dependent manner by mixed high- and low-LET radiation, and (2) selective irradiation of the microvasculature resulted in less loss of neural precursor cells than when the radiation dose was delivered uniformly to the parenchyma. This information, and in particular the approach of selectively irradiating the vasculature, may be useful in developing radioprotective compounds for use during therapeutic irradiation.     

Dosimetry of low-energy neutrons using low-pressure proportional counters

Measurements in nearly monoenergetic beams of 144, 24.5, and 2 keV neutrons and of thermal neutrons have been performed with low-pressure proportional counters. The suitability of a tissue-equivalent proportional counter (TEPC) for dosimetry of low-energy neutrons has been investigated. In contrast to higher neutron energies, the modification of the primary radiation field by the detector wall and the contribution of secondaries produced in the gas are significant. These effects have been investigated by additional measurements with a carbon-walled proportional counter. The various physical processes of neutron interaction with wall and gas of the TEPC have been analyzed, and absorbed dose, kerma, and kerma contributions from the various processes are presented. In addition, dose contributions from contaminating neutrons and photons have been obtained for the calibration fields used. The results have been related to neutron fluence. The comparison with tabulated kerma factors shows excellent agreement, indicating the suitability of the TEPC method for dosimetry of low-energy neutrons.     

Comparative study of chromosome aberration formation in lymphocytes culture under pulsed and continuous neutron irradiation

Frequencies of chromosome aberration induced by prolong (continuous) neutron radiation (dose-rate 0.17 Gy/min) and pulsed neutron radiation with ultra-high dose-rate (1-4) x 10(5) Gy/s have been studied in human blood lymphocytes at G0-stage. It was demonstrated that cytogenetic efficiency of pulsed neutrons (after the substraction of approximately 50% gamma-component from the total dose) was 2 times higher than that of continuous neutron radiation.     

Thermal and resonance neutrons generated by various electron and X-ray therapeutic beams from medical linacs installed in polish oncological centers

BackgroundHigh-energy photon and electron therapeutic beams generated in medical linear accelerators can cause the electronuclear and photonuclear reactions in which neutrons with a broad energy spectrum are produced. A low-energy component of this neutron radiation induces simple capture reactions from which various radioisotopes originate and in which the radioactivity of a linac head and various objects in the treatment room appear.AimThe aim of this paper is to present the results of the thermal/resonance neutron fluence measurements during therapeutic beam emission and exemplary spectra of gamma radiation emitted by medical linac components activated in neutron reactions for four X-ray beams and for four electron beams generated by various manufacturers’ accelerators installed in typical concrete bunkers in Polish oncological centers.Materials and methodsThe measurements of neutron fluence were performed with the use of the induced activity method, whereas the spectra of gamma radiation from decays of the resulting radioisotopes were measured by means of a portable high-purity germanium detector set for field spectroscopy.ResultsThe fluence of thermal neutrons as well as resonance neutrons connected with the emission of a 20 MV X-ray beam is ～10⁶ neutrons/cm² per 1 Gy of a dose in water at a reference depth. It is about one order of magnitude greater than that for the 15 MV X-ray beams and about two orders of magnitude greater than for the 18–22 MeV electron beams regardless of the type of an accelerator.ConclusionThe thermal as well as resonance neutron fluence depends strongly on the type and the nominal potential of a therapeutic beam. It is greater for X-ray beams than for electrons. The accelerator accessories and other large objects should not be stored in a treatment room during high-energy therapeutic beam emission to avoid their activation caused by thermal and resonance neutrons. Half-lives of the radioisotopes originating from the simple capture reaction (n,γ) (from minutes to hours) are long enough to accumulate radioactivity of components of the accelerator head. The radiation emitted by induced radioisotopes causes the additional doses to staff operating the accelerators.     

Halogenated pyrimidines as radiosensitizers for high-LET radiation

The incorporation of iododeoxyuridine (IdUrd) into Chinese hamster cells was examined as a possible radiosensitizer for fission spectrum neutrons. Dose-response curves comparing both X rays and neutrons in the same cell line with the same IdUrd replacement showed a similar radiation enhancement for IdUrd incorporation. Enhancement ratios at the 1% survival level were 1.8 for X rays and 1.5 for fission spectrum neutrons. While the mechanism of this enhancement in the response for fission neutron radiation is unclear, these positive data should support further exploration to determine if halogenated pyrimidine incorporation results in sensitization for neutron energies employed in therapy.     

Morphological changes in human melanoma cells following irradiation with thermal neutrons

Morphological changes in two human melanoma cell lines, MM96 and MM418, following irradiation with thermal neutrons, were studied using light and electron microscopy. The results show that the response of human malignant melanoma cells to neutron irradiation is both cell line dependent and dose dependent, and that in any given cell line, some cells are more resistant to irradiation than others, thus demonstrating heterogeneity in respect to radiosensitivity. Cells repopulating MM96 flasks after irradiation were morphologically similar to the cells of origin whereas in MM418 flasks cells differentiated into five morphologically distinct subgroups and showed increased melanization. The results also show that radiation causes distinctive morphological patterns of damage although ultrastructural changes unique to the high LET particles released from boron 10 neutron capture are yet to be identified.     

A review on photoneutrons characteristics in radiation therapy with high-energy photon beams

In radiation therapy with high-energy photon beams (E > 10 MeV) neutrons are generated mainly in linacs head thorough (γ,n) interactions of photons with nuclei of high atomic number materials that constitute the linac head and the beam collimation system. These neutrons affect the shielding requirements in radiation therapy rooms and also increase the out-of-field radiation dose of patients undergoing radiation therapy with high-energy photon beams. In the current review, the authors describe the factors influencing the neutron production for different medical linacs based on the performed measurements and Monte Carlo studies in the literature.     

Solid cancer risk coefficient for fast neutrons in terms of effective dose

Cancer mortality risk coefficients for neutrons have recently been assessed by a procedure that postulates for the neutrons a linear dose dependence, invokes the excess risk of the A-bomb survivors at a gamma-ray dose D(1) of 1 Gy, and assumes a neutron RBE as a function of D(1) between 20 and 50. The excess relative risk (ERR) of 0.008/mGy has been obtained for R(1) = 20 and 0.016/mGy for R(1) = 50. To compare these results to the current ICRP nominal risk coefficient for solid cancer mortality (0.045/Sv for a population of all ages; 0.036/Sv for a working population), the ERR is translated into lifetime attributable risk and is then related to effective dose. The conversion is not trivial, because the neutron effective dose has been defined by ICRP not as a weighted genuine neutron dose (neutron kerma), but as a weighted dose that includes the dose from gamma rays that are induced by neutrons in the body. If this is accounted for, the solid cancer mortality risk for a working population is found to agree with the ICRP nominal risk coefficient for neutrons in their most effective energy range, 0.2 MeV to 0.5 MeV. In radiation protection practice, there is an added level of safety, because the effective dose, E, is-for monitoring purposes-assessed in terms of the operational quantity H*, which overestimates E substantially for neutrons between 0.01 MeV and 2 MeV.     

The relationship between o.e.r., r.b.e. and number of fractions for X-ray- or neutron-induced skin damage.

The skin reactions in aerated and hypoxic mouse tails after single or fractionated doses of 250 kV X-rays or fast neutrons (6 MeV deuterons on beryllium) have been measured. The o.e.r. for one to sixteen fractions of X-rays remains constant, while that for one to ten fractions of neutrons decreases with increasing neutron fractionation and decreasing neutron dose/fraction. The o.e.r. for X-rays was 1.7, for single-neutron doses 1.4, and for ten fractions of neutrons 1.25. It was anticipated that the o.e.r. for neutron-induced damage would decrease further as neutron fractionation is increased because the contribution to damage from the highest LET components of dose, the alpha and heavy recoil particles, would increase relative to the lowe LET components. The r.b.e. values obtained for skin damage were higher at all neutron doses/fraction examined in this study on tails than all those previously obtained in studies on skin at other sites on four species. This may be due to the influence of hypoxia on the r.b.e. measurements in the mouse tail.     

High-energy neutron spectroscopy with thick silicon detectors

The high-energy neutron component of the space radiation environment in thick structures such as the International Space Station contributes to the total radiation dose received by an astronaut. Detector design constraints such as size and mass have limited the energy range of neutron spectrum measurements in orbit to about 12 MeV in Space Shuttle studies. We present a new method for high-energy neutron spectroscopy using small silicon detectors that can extend these measurements to more than 500 MeV. The methodology is based on measurement of the detector response function for high-energy neutrons and inversion of this response function with measured deposition data to deduce neutron energy spectra. We also present the results of an initial shielding study performed with the thick silicon detector system for high-energy neutrons incident on polyethylene.     

Comparison of recovery from potential mitotic abnormality in mitotically quiescent lens cells after X, neutron, and 56Fe irradiations

After exposure to various doses of 250 kVp X radiation, 0.85 Me V fission spectrum neutrons, or 600 MeV/A iron (Fe) particles, mitotically quiescent rat lens cells showed no visible evidence of radiation injury. However, following the mitogenic stimulus of wounding, mitotic abnormalities became evident when responding cells entered mitosis. Latent damage and recovery therefrom were monitored at 3, 7, 14, and 28 days after irradiation. Following doses of 1 to 10 Gy of X radiation, the recovery rate, indicated by a decrease in abnormalities with time, was proportional to dose, and the dose-effect slope decreased exponentially with time. Virtually no recovery occurred during the 28 days after 1.25 to 2.25 Gy of fission neutron radiation. After doses of 0.5 to 3.0 Gy of Fe particles, an increased expression of mitotic damage or recovery than recovery occurred. As a consequence of the differing patterns in time for expression of damage or recovery following X rays and the high-LET radiations, the relative biological effectiveness (RBE) increased from 3.6 to 16 for neutrons and from 2 to 10 for Fe particles over the 28-day observation period.     

The survival of mice and the metallothionein content of their livers and kidneys as criteria for assessing exposure to pulsed neutron radiation

The effect of pulsed neutron radiation was studied in comparison with continuous neutron radiation and continuous gamma-radiation. Animal survival and induction of metallothionein (MT) synthesis in liver and kidney of mice exposed to equivalent doses were chosen as criteria for evaluation of radiation effects. It was found that the level of MT in liver and kidney of mice exposed to neutron radiation decreased 24 hours after irradiation and then continued decreasing in kidney for 48 hours after irradiation. This is evidence of more intensive free-radical processes initiated by pulsed neutron radiation. At the same time, RBE values of pulsed neutrons did not differ significantly from that of continuous neutron radiation.     

Variation in lunar neutron dose estimates

The radiation environment on the Moon includes albedo neutrons produced by primary particles interacting with the lunar surface. In this work, HZETRN2010 is used to calculate the albedo neutron contribution to effective dose as a function of shielding thickness for four different space radiation environments and to determine to what extent various factors affect such estimates. First, albedo neutron spectra computed with HZETRN2010 are compared to Monte Carlo results in various radiation environments. Next, the impact of lunar regolith composition on the albedo neutron spectrum is examined, and the variation on effective dose caused by neutron fluence-to-effective dose conversion coefficients is studied. A methodology for computing effective dose in detailed human phantoms using HZETRN2010 is also discussed and compared. Finally, the combined variation caused by environmental models, shielding materials, shielding thickness, regolith composition and conversion coefficients on the albedo neutron contribution to effective dose is determined. It is shown that a single percentage number for characterizing the albedo neutron contribution to effective dose can be misleading. In general, the albedo neutron contribution to effective dose is found to vary between 1-32%, with the environmental model, shielding material and shielding thickness being the driving factors that determine the exact contribution. It is also shown that polyethylene or other hydrogen-rich materials may be used to mitigate the albedo neutron exposure.     

Changes in G1-phase populations in human glioblastoma and neuroblastoma cell lines influence p66/Be neutron-induced micronucleus yield

Some photon resistant tumours are sensitive to neutrons but no predictive methods exist which could identify such tumours. In a recent study addressing this clinically important issue, we demonstrated that relative biologic effectiveness (RBE) values for p(66)/Be neutrons estimated from micronucleus (MN) data correlate positively with RBE values obtained from conventional clonogenic survival data. However, not all photon-resistant cell lines showed high RBE values when the MN endpoint was used. Now, we examine how the functional status of the p53 tumour suppressor gene and radiation-induced changes in cell cycle phase populations may contribute to this discrepancy. No significant association was established between p53 status and MN yield for both photon and neutron irradiation. The data demonstrated that neutron-, but not photon-, induced MN yield is dependent on the intrinsic ability of cells to activate a G1-phase arrest. In cell lines of comparable photon sensitivity, those showing more extensive depletion of the G1 population express significantly more micronuclei per unit dose of neutrons. These results suggest that differences in cell cycle kinetics, and not the p53 status, may constitute an important factor in damage induction by high linear energy transfer (LET) irradiation and need to be considered when radiation toxicity in clinical radiobiology or radiation protection is assessed using damage endpoints.