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PURPOSE: To identify human in vitro cell lines with a high relative cellular sensitivity to fast neutrons as compared to photons and to examine their relationship to intrinsic photon radiosensitivity and cellular proliferation kinetics. METHODS AND MATERIALS: The clonogenic cell survival following exposure to low LET, 4 MeV photons or, high LET, 62.5 MeV (p-->Be+) fast neutrons and the cell kinetic parameters of 30 human in vitro cell lines, covering a wide range of histologies, were analyzed alone and with previously published data of Fertil and Malaise. The relative survival at 1.6 Gy of neutrons (SF1.6) compared to 2 Gy of photons (SF2) (the doses per fractions used in the Clatterbridge fast neutron studies) and the cell kinetic parameters of the 30 cell lines were also compared. The relative lethality of 62.5 MeV fast neutrons was assessed by comparing the ratio alpha neutrons/alpha photons to alpha photons or SF1.6 neutrons/SF2 photons to SF2 photons. Cellular proliferation kinetics were measured by flow cytometry following BrdU incorporation and the relationship of cellular proliferation to relative neutron lethality was measured by comparing the alpha neutron/alpha photon ratio to the labelling index (LI), potential doubling (Tpot) and ploidy. RESULTS: The majority of cell survival curves obtained following exposure to 62.5 MeV fast neutrons were curvilinear with beta values of similar order to those obtained with low LET 4 MeV photons. Comparison of alpha values for neutrons and photons revealed a relatively neutron sensitive subset of 9 out of 30 in vitro cell lines. This subset was not, however, distinguishable when 1.6 Gy of neutrons was compared to 2 Gy of photons. There was no correlation between cell survival with neutrons or photons and the cell kinetic parameters Tpot or LI or with DNA ploidy. CONCLUSIONS: The use of in vitro assays of neutron and photon radiosensitivity irrespective of cell kinetic parameters allows identification of neutron sensitive cell populations when the ratio of the alpha values for neutrons and photons is compared to the reciprocal of the alpha photon value. This relationship is not apparent when fractions of 2 Gy of photons are compared to 1.6 Gy of neutrons. Whether or not this identification can be borne out in fractionated regimes in the clinic remains to be proved.
The frontal collisions of a laser beam with relativistic electrons result in Compton-backscattered photons. The energy of these photons is dependent on the laser and electron energy in the range from kilo-electron-volts to tens of mega-electron-volts. In a sufficiently narrow backscattering angle the photons are nearly monochromatic. Over the past 30 years there have been several attempts to produce photon beams by laser backscattering from relativistic electrons stored in magnetic ring structures. One aim is to produce photons in the high mega-electron-volt energy range with fluxes useful for nuclear physics research; another is to produce photons in the high kilo-electron-volt energy range, which would be useful for medical applications, such as coronary angiography or treatment of tumour. Our present interest is to investigate the possibility of using 34 keV to 10 MeV photon beams for applications in stereotactic functional radiosurgery. We foresee the possibility of neurosurgical operations through the intact skull with precise and effective destruction of deeply lying millimetre-sized targets with minimal effects on intervening structures, high reproducibility and good prediction of the results. Our paper presents: a Monte Carlo study of radiosurgery based on cross firing with 34 keV to 100 MeV photon beams and 200 and 580 MeV proton beams, a theoretical description of the kinematics of Compton backscattering and estimates of the backscattered photon flux from several combinations of laser cavities at Nd:YAG (1.17 eV) and CO2 (0.117 eV) laser energies and electron storage rings energies in the range 0.1-1.3 GeV. As examples, existing magnetic structures, such as the DA phi NE Accumulator in the lower energy range and the Trieste Synchrotron Light Source ELETTRA in the higher energy range have been utilized in the calculations. The Monte Carlo study has shown that radiosurgery with photon beams of energies in mega-electron-volt energy range enables precise destruction of deeply lying millimetre-sized targets with minimal effects on intervening structures. Its precision is comparable to that of radiosurgery with 200-580 MeV proton beam, but our hope is that radiosurgery with lower energy photon beams could be more precise and less expensive. An average dose of 200 Gy can be delivered to a target of diameter 2 mm at the centre of an 18 cm diameter phantom in 1 h using photon beams of fluences 7.3 x 10(10), 1.8 x 10(10), 6.5 x 10(8), 2.2 x 10(8), 8.6 x 10(7) and 7.8 x 10(6) photons per second at 34 keV, 100 keV, 1 MeV, 3 MeV, 10 MeV and 100 MeV per cross section of beam of 2 mm diameter, respectively. 34-100 keV photon beams were studied in the hope of finding a strong enhancement of their efficiency if a stable high-Z element were to be introduced into the target's DNA. It is shown that, with a low-energy ring running at about 0.4 GeV and a Nd:YAG laser, it would be possible to obtain the required 3 MeV photon beam flux to deliver the average dose within 1 h, assuming an average distance between the source and the target of about 5 m. With a similar machine used at about 1.3 GeV and a CO2 laser, a 3 MeV photon beam is obtained and the exposure time can be reduced to less than 1 min, assuming a roughly 10 m distance between source and target (here a beam angle of 0.1 mrad only had to be considered due to the larger angular energy and yield spread). With a lower electron energy of 138 MeV and a CO2 laser, a 34 keV photon beam can be produced. More than 45 h would be needed to deliver the same dose. We hope that this time could be shortened considerably if stable iodine were introduced into the target with the help of a DNA-seeking molecular carrier. In this case the geometrical precision would be further improved.
We report the calcium-dependent emission spectral properties of the calcium probe Indo-1 for three-photon excitation. We found that Indo-1 could be readily excited with the femtosecond pulses from a mode-locked Ti:sapphire laser at 885 nm. This wavelength is too long for two-photon excitation, which is expected to occur for wavelengths no longer than twice the longest single-photon absorption wavelength of 400 nm. For excitation at 885 nm the emission intensity was found to depend on the cube of the laser power, as expected for simultaneous interaction with three photons. At wavelengths below 840 nm the emission intensity depends on the square of the laser power, indicating two-photon excitation at shorter wavelengths. The intensity decays of Indo-1 were found to be dependent on Ca2+ and essentially identical for one- and three-photon excitation. The emission anisotropy of Indo-1 was found to be considerably higher for three-photon excitation than for one-photon excitation, consistent with cos6 theta photoselection, as compared with cos2 theta photoselection for one-photon excitation. The high values of the anisotropy are in agreement with those expected for a three-photon process. Calcium-dependent emission spectra were observed for Indo-1 with three-photon excitation, demonstrating that three-photon excitation of Indo-1 can be used for calcium imaging by emission intensity ratio measurements. The calcium-dependent emission spectra indicate a higher three-photon cross-section for the calcium-free form of Indo-1 than for the calcium-bound form. The possible advantages of three-photon excitation include the availability of the appropriate wavelengths with solid-state lasers, enhanced spatial resolution due to a reduced size of the excited volume, absence of light quenching, and possibly high selectivity of the three-photon excitation process.
Photon counting x-ray imaging provides efficient rejection of the electronics noise, no pulse height (Swank) noise, less noise due to optimal photon energy weighting and the possibility of energy resolved image acquisition. These advantages apply also to CT when projection data are acquired using a photon counting detector. However, photon counting detectors assign a weighting factor of 1 to all detected photons whereas the weighting factor of a charge integrating detector is proportional to the energy of the detected photon. Therefore, data collected by photon counting and charge integrating detectors represent the 'hardening' of the photon beam passed through the object differently. This affects the beam hardening artefacts in the reconstructed CT images. This work represents the first comparative evaluation of the effect of photon counting, charge integrating and energy weighting photon detectors on beam hardening artefacts in CT. Beam hardening artefacts in CT images were evaluated for 20 cm and 14 cm diameter water cylinders with bone and low contrast inserts, at 120 kVp and 90 kVp x-ray tube voltages, respectively. It was shown that charge integrating results in 1.8% less beam hardening artefacts from bone inserts (i.e., CT numbers in the 'shadow' of the bone are less by 1.8% as compared to CT numbers over the periphery of the image), as compared to photon counting. However, optimal photon energy weighting, which provides highest SNR, results in 7.7% higher beam hardening artefacts from bone inserts as compared to photon counting. The magnitude of the 'cupping' artefacts was lower by 1% for charge integrating and higher by 6.1% for energy weighting acquisitions as compared to photon counting. Only the photon counting systems provide an accurate representation of the beam hardening effect due to its flat energy weighting. Because of their energy dependent weighting factors, the charge integrating and energy weighting systems do not provide accurate representation of the beam hardening effect.
Photon dose calculation algorithms (such as the pencil beam and collapsed cone, CC) model the attenuation of a primary photon beam in media other than water, by using pathlength scaling based on the relative mass density of the media to water. In this study, we assess if differences in the electron density between the water and media, with different atomic composition, can influence the accuracy of conventional photon dose calculations algorithms. A comparison is performed between an electron-density scaling method and the standard mass-density scaling method for (i) tissues present in the human body (such as bone, muscle, etc.), and for (ii) water-equivalent plastics, used in radiotherapy dosimetry and quality assurance. We demonstrate that the important material property that should be taken into account by photon dose algorithms is the electron density, and not the mass density. The mass-density scaling method is shown to overestimate, relative to electrondensity predictions, the primary photon fluence for tissues in the human body and water-equivalent plastics, where 6%-7% and 10% differences were observed respectively for bone and air. However, in the case of patients, differences are expected to be smaller due to the large complexity of a treatment plan and of the patient anatomy and atomic composition and of the smaller thickness of bone/air that incident photon beams of a treatment plan may have to traverse. Differences have also been observed for conventional dose algorithms, such as CC, where an overestimate of the lung dose occurs, when irradiating lung tumors. The incorrect lung dose can be attributed to the.incorrect modeling of the photon beam attenuation through the rib cage (thickness of 2-3 cm in bone upstream of the lung tumor) and through the lung and the oversimplified modeling of electron transport in convolution algorithms. In the present study, the overestimation of the primary photon fluence, using the mass-density scaling method, was shown to be a consequence of the differences in the hydrogen content between the various media studied and water. On the other hand, the electron-density scaling method was shown to predict primary photon fluence in media other than water to within 1%-2% for all the materials studied and for energies up to 5 MeV. For energies above 5 MeV, the accuracy of the electron-density scaling method was shown to depend on the photon energy, where for materials with a high content of calcium (such as bone, cortical bone) or for primary photon energies above 10 MeV, the pair-production process could no longer be neglected. The electron-density scaling method was extended to account for pair-production attenuation of the primary photons. Therefore the scaling of the dose distributions in media other than water became dependent on the photon energy. The extended electron-scaling method was shown to estimate the photon range to within 1% for all materials studied and for energies from 100 keV to 20 MeV, allowing it to be used to scale dose distributions to media other than water and generated by clinical radiotherapy photon beams with accelerator energies from 4 to 20 MV.
A realistic model of photon beams generated by clinical linear accelerators has been incorporated in a convolution/superposition method to compute dose distributions in photon treatment fields. In this beam model, a primary photon source represents photons directly from the target, and an extra-focal photon source represents scattered photons from the primary collimator and the flattening filter. Monte Carlo simulation was used to study clinical linear accelerators producing photon beams. From the output of the Monte Carlo simulation, the fluence and spectral distributions of each photon component, as well as the geometrical characteristics of each photon source with respect to its distance to the isocenter and its source distribution, were analyzed. These quantities were used to reproduce realistic photon distributions in treatment fields, and thus to compute dose distributions using the convolution method. Our results showed that compared to the primary photon fluence, the extra-focal photon fluence from the primary collimator and the flattening filter was 11%-16% at the isocenter, among which 70% was contributed by the flattening filter. The variation of extra-focal photons in different treatment fields was predicted accurately by accounting for the finite size of the extra-focal source. Compared to measurements, dose distributions in photon treatment fields, including those of asymmetric jaw settings and at different SSDs were calculated accurately, particularly in the penumbral region, by using the convolution method with the new dual source photon beam model.
Two-photon fluorescence excitation has been found to be a very powerful method for enhancing the sensitivity and resolution in far-field light microscopy. Two-photon fluorescence excitation also provides a substantially background-free detection on the single-molecule level. It allows direct monitoring of formation of labelled biomolecule complexes in solution. Two-photon excitation is created when, by focusing an intensive light source, the density of photons per unit volume and per unit time becomes high enough for two photons to be absorbed into the same chromophore. In this case, the absorbed energy is the sum of the energies of the two photons. In two-photon excitation, dye molecules are excited only when both photons are absorbed simultaneously. The probability of absorption of two photons is equal to the product of probability distributions of absorption of the single photons. The emission of two photons is thus a quadratic process with respect to illumination intensity. Thus in two-photon excitation, only the fluorescence that is formed in the clearly restricted three-dimensional vicinity of the focal point is excited. We have developed an assay concept that is able to distinguish optically between the signal emitted from a microparticle in the focal point of the laser beam, and the signal emitted from the surrounding free labelled reagent. Moreover, the free labels outside the focal volume do not contribute any significant signal. This means that the assay is separation-free. The method based on two-photon fluorescence excitation makes possible fast single-step and separation-free immunoassays, for example, for whole blood samples. Since the method allows a separation-free assay in very small volumes, the method is very useful for high-throughput screening assays. Consequently we believe that two-photon fluorescence excitation will make a remarkable impact as a research tool and a routine method in many fields of analysis.
Fluorescence correlation spectroscopy (FCS) is generally used to obtain information about the number of fluorescent particles in a small volume and the diffusion coefficient from the autocorrelation function of the fluorescence signal. Here we demonstrate that photon counting histogram (PCH) analysis constitutes a novel tool for extracting quantities from fluorescence fluctuation data, i.e., the measured photon counts per molecule and the average number of molecules within the observation volume. The photon counting histogram of fluorescence fluctuation experiments, in which few molecules are present in the excitation volume, exhibits a super-Poissonian behavior. The additional broadening of the PCH compared to a Poisson distribution is due to fluorescence intensity fluctuations. For diffusing particles these intensity fluctuations are caused by an inhomogeneous excitation profile and the fluctuations in the number of particles in the observation volume. The quantitative relationship between the detected photon counts and the fluorescence intensity reaching the detector is given by Mandel's formula. Based on this equation and considering the fluorescence intensity distribution in the two-photon excitation volume, a theoretical expression for the PCH as a function of the number of molecules in the excitation volume is derived. For a single molecular species two parameters are sufficient to characterize the histogram completely, namely the average number of molecules within the observation volume and the detected photon counts per molecule per sampling time epsilon. The PCH for multiple molecular species, on the other hand, is generated by successively convoluting the photon counting distribution of each species with the others. The influence of the excitation profile upon the photon counting statistics for two relevant point spread functions (PSFs), the three-dimensional Gaussian PSF conventionally employed in confocal detection and the square of the Gaussian-Lorentzian PSF for two photon excitation, is explicitly treated. Measured photon counting distributions obtained with a two-photon excitation source agree, within experimental error with the theoretical PCHs calculated for the square of a Gaussian-Lorentzian beam profile. We demonstrate and discuss the influence of the average number of particles within the observation volume and the detected photon counts per molecule per sampling interval upon the super-Poissonian character of the photon counting distribution.
PURPOSE: To determine the incidence of acute toxicity and treatment interruption associated with electron and photon craniospinal irradiation (CSI) in children treated with or without chemotherapy. MATERIALS AND METHODS: A retrospective study involving a computerized search of the radiotherapy database at the University of Texas M. D. Anderson Cancer Center identified a total of 79 eligible patients </=18 years old who had received electron (n = 46) or photon (n = 33) CSI from October 1980 to March 2000. Acute toxicity was graded according to the 1998 National Cancer Institute Common Toxicity Criteria. Chemotherapy sequencing was categorized as before or after CSI or no chemotherapy. The incidences of weight loss and skin toxicity were recorded and differences in treatment interruption and hematologic values with respect to modality used (electron vs. photon), age (</=6 or >6 years), and sequencing of chemotherapy were compared using chi-square tests. RESULTS: The median age of the electron group was lower than that of the photon group (6.7 years and 11.7 years, respectively). The two groups were otherwise well matched in terms of median spinal dose (31.1 vs. 33.3 Gy), fraction size (1.57 vs. 1.63 Gy), and total treatment time (32.4 vs. 30.7 days). Only 2 patients in each group (photon and electron) had a treatment break (>3 days). The mean number of days interrupted was 0.94 (photon) and 1.1 (electron) (p = 0.72). The electron and photon groups were well balanced in terms of receiving pre-CSI chemotherapy (37% vs. 41%, p = 0.776). Chemotherapy given before radiotherapy vs. after or not at all was associated with an increased incidence of Grade 3-4 leukopenia (76% vs. 49%, p = 0.02), thrombocytopenia (90% vs. 10%, p = 0), and neutropenia (50% vs. 15%, p = 0.005). A younger age was associated with Grade 3-4 thrombocytopenia (29% vs. 8.7%, p = 0.034), and decreased hemoglobin (29% vs. 6.5%, p = 0.014). The incidence of leukocyte depression of Grade 3-4 toxicity was 62% in the electron group and 32% in the photon group (p = 0.018). The incidence of Grade 3-4 platelet toxicity was higher with electrons (21%) than with photons (4%), but the difference was of borderline significance (p = 0.053). The difference in Grade 1-2 weight loss was not statistically significant (86% electron vs. 55% photon groups, p = 0.53), and Grade 3-4 weight loss did not occur within the entire study group. Most patients experienced Grade 0-1 radiation dermatitis with either electrons or photons. CONCLUSION: A younger patient age (</=6 years) and the sequence of chemotherapy before radiotherapy were associated with increased acute hematologic toxicity. Grade 3-4 weight loss, a surrogate for gastrointestinal toxicity, was not observed within the entire population. Despite the increased acute hematologic toxicities associated with CSI in very young children, the vast majority of patients were able to complete photon or electron CSI with minimal to no treatment interruptions. CSI with the electron technique can be used as an alternative to photon CSI in young children, and in our experience does not require the routine use of hematopoietic growth factors.
6MAP is a fluorescent analogue of adenine that undergoes Watson-Crick base pairing and base stacking in double-stranded DNA. The one-photon absorption spectrum of 6MAP is characterized by a maximum around 330 nm with moderate quantum yield fluorescence centered at about 420 nm. To take advantage of this probe for confocal and single-molecule microscopy, it would be advantageous to be able to excite the analogue via two photons. We report the first determination of the two-photon excitation cross section and spectrum for 6MAP from 614 to 700 nm. The power dependence of the fluorescence indicates that emission results from the absorption of two photons. The one-photon and two-photon emission line shapes are identical within experimental error. A study of the concentration dependence of the fluorescence yield for one-photon excitation shows no measurable quenching up to about 5 microM. The maximum in the two-photon excitation spectrum gives a two-photon cross section, delta(TPE), of 3.4 +/- 0.1 Goeppert-Mayer (G.M.) at 659 nm, which correlates well with the one-photon absorption maximum. This compares quite favorably with cross sections of various naturally fluorescent biological molecules such as flavins and nicotiamide. In addition, we have also obtained the two-photon-induced fluorescence emission spectrum of quinine sulfate. It is approximately the same as that for one-photon excitation, suggesting that two-photon excitation of quinine sulfate may be used for calibration purposes.
Dual energy photon irradiation (6 MV and 20 MV) was compared to conventional treatment planning with 6 MV photons in a lateralized intracranial malignancy. Dose volume analysis was performed of both the tumor plus a 2 cm margin (target volume, TV) and normal tissues (NT). Parallel opposed treatment using weightings of 1:1, 1.5:1, and 2:1 were compared for 6 MV photons alone or in combination with 20 MV photons. Uniform treatment of the TV was accomplished within the 60 Gy isodose. Significant differences were observed, however, in NT volumes receiving greater than or equal to 60 Gy and 45-59 Gy. Dual photon energy reduced treatment of NT volumes to greater than or equal to 60 Gy by 13% (177 cm3 vs 204 cm3 in 2:1 weighting) to 70% (147 cm3 vs 498 cm3 in 1:1 weighting) for comparable plans. Dose optimization was also performed for both 6 MV alone or in combination with 20 MV photons. Usual approaches to achieve dose lateralization with conventional isocentric techniques were applied including parallel opposed 6 MV photons ipsilaterally weighted 3.4:1 (POP), and a 110 degrees arc rotational field used to limit treatment to the eye (ARC). Dual energy photon optimized plans included a three beam parallel opposed plan (TOP) and a mixed photon ipsilateral (IPSI) approach. The technique using parallel opposed 20 MV photons and ipsilateral 6 MV photons (TOP) used beam weightings of 1.1 (contralateral 20 MVX): 1.6 (ipsilateral 6 MVX): 1 (ipsilateral 20 MVX) to achieve dose optimization. The ipsilateral approach with 6 MVX and 20 MVX (IPSI) used beam weightings of 1:1.4, respectively. All optimized plans demonstrated a 41% (120 cm3; POP) to 53% (95 cm3; TOP) improvement over parallel opposed 6 MV photons weighted 2:1 (204 cm3) in NT volume receiving greater than or equal to 60 Gy. Comparison of optimized treatment showed the IPSI plan to be superior, treating 12% of NT volume to greater than or equal to 60 Gy and 38% to 45-59 Gy; the 6 MV POP plan resulted in NT volumes of 15% and 51%, respectively, for those dose levels. Dual photon energy irradiation of lateralized intracranial malignancies allows reduction of dose to normal tissue volumes while achieving excellent coverage of the target volume. Treatment planning should be performed in all lateralized intracranial lesions to achieve dose optimization exploiting depth dose characteristics.
A vectorized Monte Carlo computer code has been developed for modeling photon transport in single photon emission computed tomography (SPECT). The code models photon transport in a uniform attenuating region and photon detection by a gamma camera. It is adapted from a history-based Monte Carlo code in which photon history data are stored in scalar variables and photon histories are computed sequentially. The vectorized code is written in FORTRAN77 and uses an event-based algorithm in which photon history data are stored in arrays and photon history computations are performed within DO loops. The indices of the DO loops range over the number of photon histories, and these loops may take advantage of the vector processing unit of our Stellar GS1000 computer for pipelined computations. Without the use of the vector processor the event-based code is faster than the history-based code because of numerical optimization performed during conversion to the event-based algorithm. When only the detection of unscattered photons is modeled, the event-based code executes 5.1 times faster with the use of the vector processor than without; when the detection of scattered and unscattered photons is modeled the speed increase is a factor of 2.9. Vectorization is a valuable way to increase the performance of Monte Carlo code for modeling photon transport in SPECT.