Description of an XRF system for multielemental analysis

An X-ray fluorescence (XRF) system that uses radioisotopes in an orthogonal configuration between the source, sample, and detector is described. The advantage of such a system is that for large (bulk) samples or in vivo measurements, the background caused by Compton scattering in the sample is minimized. High reproducibility for nonuniform samples is obtained by reducing the sample size and thus the effects of nonuniformity in the spatial response of such a system. Germane to any accurate analytical method is the use of proper mathematical algorithms for data evaluation. The problem is acute, in particular, when photopeaks with low counting statistics are to be analyzed. In the case of a single photopeak on flat background, optimal energy window size, which maximizes the signal-to-noise ratio, for trapezoidal intergration is described. The sensitivity and minimum detection limit at different energies together with background considerations are discussed.     

An APD-based iterative reconstruction method for simultaneous technetium-99m/iodine-123 SPECT imaging

Dual-isotope SPECT (DI-SPECT) studies offer significant advantages over sequential scans, foremost among them faster acquisition and perfect image registration. However, reconstructed images may be affected by substantial cross-talk contamination rendering them inadequate for diagnosis. This effect is especially strong for isotopes with close photopeak energies, such as ^99mTc (140 keV) and ¹²³I (159 keV). In this paper we present an iterative DI-SPECT reconstruction method which includes accurate, analytically computed scatter corrections provided by the APD (analytical photon distribution) algorithm. This algorithm calculates first and second order Compton scatter (based on the Klein–Nishina formula) and first order Rayleigh scatter. Both self-scatter and cross-talk between the two isotopes are evaluated using patient specific attenuation maps and an initial activity distribution estimate. To validate our method we performed experiments using the Data Spectrum, Inc. thorax phantom and a SPECT/CT camera system. Reconstructed images demonstrate significant improvement in data quantitation. Their quantitative accuracy increases up to a factor of two, even for activity ratios which strongly enhance cross-talk effects and seriously degrade projections.     

Effect of number of views on cross-sectional Compton imaging: A fundamental study with backprojection

IntroductionWe have been developing a medical imaging technique using a Compton camera, which is expected to reconstruct three-dimensional images. If the number of views is not sufficient, star-shaped artifacts (streak artifacts) could arise in cross-sectional images. Therefore, we estimated the point spread function (PSF) of cross-sectional Compton images and the effect of the number of views by Monte Carlo simulations and experimental studies.Materials and methodsA cross-sectional Compton image was reconstructed using a dataset comprising 719 view directions and PSF was analyzed using a radial distribution. The peak height, full width at half maximum (FWHM), background (BG), and residual sum of squares (RSS) were calculated from the obtained PSF. In addition, RSSs were plotted against the number of views to estimate the required number to suppress star-shaped artifacts.ResultsThere was no correlation found between the number of views and both FWHM (16 mm) and peak/BG ratio (∼1 × 10⁴). RSSs were reduced with the number of views and approached the minimum asymptotically. Correlation was observed between the required number of views and the number of Compton events used for image reconstruction.ConclusionWe determined the PSF of cross-sectional Compton images and the effect of the number of views on the images. The required number of views to suppress the star-shaped artifact is related to the square root of the number of Compton events used to reconstruct the image. From this study, we concluded that 21 or more views are required for clinical purposes.     

The Bethe surface of liquid water

The Bethe surface of liquid water, which was previously calculated by using a semiempirical model, is compared with recently available data from IXS experiments (inelastic x-ray scattering; Compton scattering of high energy photons) in liquid water. No alarming discrepancy is found for a global view of the Bethe surface, in part because the two sets of data have been constrained by the Bethe sum rule. The shape of the Bethe ridge given by the new data is broader than that obtained through the impulse approximation. The extrapolation to the optical limit (viz., at zero momentum transfer) is shown, and the reliability of these data is discussed in detail. Received: 11 September 1998 / Accepted in revised form: 15 February 1999     

Analysis of self-associating proteins by singular value decomposition of solution scattering data

We describe a method by which a single experiment can reveal both association model (pathway and constants) and low-resolution structures of a self-associating system. Small-angle scattering data are collected from solutions at a range of concentrations. These scattering data curves are mass-weighted linear combinations of the scattering from each oligomer. Singular value decomposition of the data yields a set of basis vectors from which the scattering curve for each oligomer is reconstructed using coefficients that depend on the association model. A search identifies the association pathway and constants that provide the best agreement between reconstructed and observed data. Using simulated data with realistic noise, our method finds the correct pathway and association constants. Depending on the simulation parameters, reconstructed curves for each oligomer differ from the ideal by 0.05-0.99% in median absolute relative deviation. The reconstructed scattering curves are fundamental to further analysis, including interatomic distance distribution calculation and low-resolution ab initio shape reconstruction of each oligomer in solution. This method can be applied to x-ray or neutron scattering data from small angles to moderate (or higher) resolution. Data can be taken under physiological conditions, or particular conditions (e.g., temperature) can be varied to extract fundamental association parameters (ΔH_ass, ΔS_ass).     

Sarcoptes scabiei var. hominis: Three-dimensional structure of a female imago and crusted scabies lesions by X-ray micro-CT

The three-dimensional structure of scabies mites (Sarcoptes scabiei var. hominis) and keratin layers affected by crusted scabies lesions were obtained using X-ray computed tomography at sub-micrometer and micrometer resolution, respectively (X-ray micro-CT). Clear three-dimensional images including internal structure of scabies mites were obtained. Utilizing reconstructed micro-CT data, the sections of the capitulum (head part), digestive organs, and legs are shown. The reconstructed capitulum shows a jaw-like structure capable of penetrating the keratin layer of the skin. The tip of the forelegs of female scabies mites has a flat disk structure that may be used to grasp the skin surface. The keratin layer of a crusted scabies lesion spontaneously exfoliated from a patient was also reconstructed by the X-ray micro-CT technique. Extracted sections from CT data revealed a network structure of tunnels made by scabies mites with numerous larvae and eggs inside the tunnels.     

A methodology for estimating the shape of biconcave red blood cells using multicolor scattering images

In this paper, a novel methodology for estimating the shape of human biconcave red blood cells (RBCs), using color scattering images, is presented. The information retrieval process includes, image normalization, features extraction using two-dimensional discrete transforms, such as angular radial transform (ART), Zernike moments and Gabor filters bank and features dimension reduction using both independent component analysis (ICA) and principal component analysis (PCA). A radial basis neural network (RBF-NN) estimates the RBC geometrical properties. The proposed method is evaluated in both regression and identification tasks by processing images of a simulated device used to acquire scattering phenomena of moving RBCs. The simulated device consists of a tricolor light source (light emitting diode – LED) and moving RBCs in a thin glass. The evaluation database includes 23,625 scattering images, obtained by means of the boundary element method. The regression and identification accuracy of the actual RBC shape is estimated using three feature sets in the presence of additive white Gaussian noise from 60 to 10 dB SNR and systematic distortion, giving a mean error rate less than 1% of the actual RBC shape, and more than 99% mean identification rate.     

Ultrasound Elastography of the Prostate Using an Unconstrained Modulus Reconstruction Technique: A Pilot Clinical Study

A novel full-inversion-based technique for quantitative ultrasound elastography was investigated in a pilot clinical study on five patients for non-invasive detection and localization of prostate cancer and quantification of its extent. Conventional-frequency ultrasound images and radiofrequency (RF) data (~5 MHz) were collected during mechanical stimulation of the prostate using a transrectal ultrasound probe. Pre and post-compression RF data were used to construct the strain images. The Young's modulus (YM) images were subsequently reconstructed using the derived strain images and the stress distribution estimated iteratively using finite element (FE) analysis. Tumor regions determined based on the reconstructed YM images were compared to whole-mount histopathology images of radical prostatectomy specimens. Results indicated that tumors were significantly stiffer than the surrounding tissue, demonstrating a relative YM of 2.5 ± 0.8 compared to normal prostate tissue. The YM images had a good agreement with the histopathology images in terms of tumor location within the prostate. On average, 76% ± 28% of tumor regions detected based on the proposed method were inside respective tumor areas identified in the histopathology images. Results of a linear regression analysis demonstrated a good correlation between the disease extents estimated using the reconstructed YM images and those determined from whole-mount histopathology images (r² = 0.71). This pilot study demonstrates that the proposed method has a good potential for detection, localization and quantification of prostate cancer. The method can potentially be used for prostate needle biopsy guidance with the aim of decreasing the number of needle biopsies. The proposed technique utilizes conventional ultrasound imaging system only while no additional hardware attachment is required for mechanical stimulation or data acquisition. Therefore, the technique may be regarded as a non-invasive, low cost and potentially widely-available clinical tool for prostate cancer diagnosis.     

Individual filamentous phage imaged by electron holography

An in-line electron hologram of an individual f1.K phage was recorded with a purpose-built low energy electron point source (LEEPS) microscope. Cryo-microscopic methods were employed to prepare the specimen so that a single phage could be presented to the coherent low energy electrons: An aqueous phage suspension was applied to a thin carbon membrane with micro-machined slits. The membrane was rapidly cooled to freeze the remaining water as an amorphous ice sheet, which was then sublimated at low temperatures and pressures to leave individual free-standing phages suspended across slits. An image of a phage particle, depicted as the amplitude of the object wave, was reconstructed numerically from a digitized record of the hologram, obtained using 88 eV coherent electrons. The reconstructed image shows a single phage suspended across a slit in a supporting carbon membrane, magnified by a factor of 100,000. The width and shape in the reconstructed image compared well with a TEM image of the same filament. It is thus possible to record and reconstruct electron holograms of an individual phage. The challenge now is to improve the resolution of reconstructed images obtained by this method and to extend these structural studies to other biological molecules.     

Antimony accumulation and antioxidative responses in four fern plants

Antimony (Sb) toxicity and contamination has become a growing concern in recent years. Remediation of Sb contamination using plants may be an effective approach. This study aimed to investigate the potential of antimony (Sb) tolerance and accumulation by plants, as well as to understand the antioxidative responses to Sb. One set of hydroponic trials was set up using four species of fern plants, including Pteris cretica (PCA), Microlepia hancei (MH), Cyrtomium fortunei (CYF) and Cyclosorus dentatus (CYD). Ferns were grown for 2 weeks in nutrient solution containing a medium (5 mg L^?1) and a high (20 mg L^?1) rate of Sb, with no Sb added as the control. The biomass of fern PCA remained constant with Sb addition, whereas the biomass of ferns CYF, MH and CYD at the high Sb rate exposure decreased by 12.5%, 35.0% and 38.3%, respectively as compared with their controls. This suggests a high to low Sb tolerance order for these four fern plants. For all of these fern plants, more Sb was accumulated in the roots than in the fronds. Antimony concentration in the roots at the high rate of Sb addition was recorded, on average, as 358 mg kg^?1 for fern PCA, 224 mg kg^?1 for fern CYF, 124 mg kg^?1 for fern CYD and 123 mg kg^?1 for fern MH. A high rate of addition of Sb increased the contents of malondialdehyde (MDA) by 41.3% and 171.6% for ferns MH and CYD, respectively, as compared with their controls. No changes for MDA contents were observed in ferns PCA and CYF with Sb addition, indicating no lipid peroxidation reaction in these two plants. At a medium rate of Sb addition, the activities of peroxidase, catalase and ascorbate peroxidase in fern PCA were much higher than those in ferns CYF, CYD and MH, demonstrating the important role of these three enzymes in resisting Sb toxicity. The consistency in unchanged biomass, high accumulation of Sb in roots, lower MDA contents, as well as high enzyme production in fronds, indicated that fern PCA was more tolerant to Sb than the other three fern plants. Antioxidative enzymes (peroxidase, catalase and ascorbate peroxidase) might be involved in Sb toxicity resistance of fern PCA.     

Comparison of Contrast-Enhanced Isotropic 3D-GRE-T1WI Sequence versus Conventional Non-Isotropic Sequence on Preoperative Staging of Cervical Cancer

Purpose

To compare contrast-enhanced isotropic 3D-GRE-T1WI sequence vs. conventional non-isotropic sequence in terms of image quality, estimated signal-to-noise ratio (eSNR), relative tumor contrast and performance of cervical cancer staging.

Methods

This retrospective study was approved by the institutional review board, and informed consent was waived. Seventy-one patients (47 ± 9.4 years), with pathologically-confirmed cervical cancer underwent axial contrast-enhanced 1mm³ isotropic 3D-GRE-T1WI sequence (herein referred to Isotropy), and 3-mm-thick non-isotropic sagittal and coronal sequences. Image quality score, eSNR and relative contrast between tumor to myometrium, gluteal muscle, and fat respectively, were compared between 3-mm-thick reconstructed images from Isotropy and directly scanned non-isotropic images by paired t-test. Difference in tumor staging obtained from Isotropy and combined Three-planes including reconstructed axial images, directly scanned sagittal and coronal sequence were compared by McNemar test.

Results

Both sequences showed similar image quality. Reconstructed images demonstrated higher eSNR, equal or lower relative tumor contrast compared with non-isotropic images. Compared with performing diagnosis on Three-planes, both reviewers showed higher accuracy when diagnosing vaginal invasion on Isotropy (p = 0.039 and 0.003, respectively).

Conclusion

Compared with non-isotropic sequence, 3.0T MR isotropic 3D-GRE-T1WI sequence exhibited better eSNR, providing more reliable clinical information for preoperative staging of cervical cancer.     

Toward a dedicated system for quantitative SPECT mammotomography

This work extends our previous quantitative brain SPECT research to breast imaging-SPECT mammotomography. A cost-effective dedicated SPECT mammotomography system is presented, which aims to acquire sufficient information for efficient reconstruction of large volumetric images. A very short focal-length fan-beam collimation is designed to maximize the information collected from a relitively small vital organ that has a uniform attenuation property. Data noise of a Poisson nature is accurately modeled and effectively treated in sinogram space, followed by efficient compensation for Compton scattering, uniform attenuation, and collimater response. This sinogram-space statistical approach has the potential to reconstruct high-resolution images at a very fast speed. Yet it provides the same image quality as iterative reconstruction algorithms did, in terms of abnormality detectability by observer studies.     

Imaging of multi-color fluorescence emission from leaf tissues

Multi-color fluorescence emission from leaf tissues is presented as a powerful reporter on plant biochemistry and physiology that can be applied both at macro- and micro-scales. The blue–green fluorescence emission is typically excited by ultraviolet (UV) excitation. However, this approach cannot be applied in investigating intact leaf interior because the UV photons are largely absorbed in the epidermis of the leaf surface. This methodological barrier is eliminated by replacing the UV photon excitation by excitation with two infra-red photons of the same total energy. We demonstrate this approach by using two-photon excitation for microscopy of Arabidopsis thaliana leaves infected by pathogenic bacterium Pseudomonas syringae. The leaf structures are visualized by red chlorophyll fluorescence emission reconstructed in 3-D images while the bacteria are detected by the green emission of engineered fluorescence protein.     

A comparison of photographic analyses used to quantify zooxanthella density and pigment concentrations in Cnidarians

Many cnidarians exist in an obligatory mutualism with dinoflagellates commonly called zooxanthellae. When these symbioses are stressed, zooxanthella densities often decrease (i.e., bleaching), resulting in reduced host fitness or mortality. Because zooxanthellae play a prominent role in the coloration of hosts, several analyses of reflected spectra from photographic images have been developed to quantify zooxanthella densities and serve as a proxy for invasive sampling methods. To date these techniques have not been compared. In this study, global information system (GIS) tools, commonly used with aerial and satellite images, and photographs of healthy and bleached sea anemones, Aiptasia pallida (Verrill), were used to compare these image analysis methods. Zooxanthella densities and chlorophyll-a concentrations were correlated with image brightness (i.e., digital number) in: the red, green, and blue bands (RGB); the average of the three RGB bands (RGB/3); intensity and saturation bands (IHS); and using a principal components analysis (PCA) of the RGB bands. RGB brightness correlations with zooxanthella densities and chlorophyll-a concentrations were highest using the blue band, followed by green, then red. Using any one band within RGB, however, restricts comparisons to similar color morphs. RGB/3, IHS or PCA transformations enable intra and inter-specific comparisons where colors may vary. Among these transformations, PCA and intensity had higher correlations, followed by RGB/3, then saturation. RGB/3 and IHS, unlike PCA, ignore the correlations between the three RGB bands, treating each pixel independently. PCA uses these correlations, and in doing so lessens the effects of heteroscedasticity in the data. In addition, the observed reciprocal relationship of intensity and saturation may serve as a standardized criterion for bleaching. Finally, this study demonstrates that GIS has broad interdisciplinary applications for spatial and spectral analyses from the individual colony to reef scale assessments.     

Reprint of "Three-dimensional elemental mapping of phosphorus by quantitative electron spectroscopic tomography (QuEST)" [J. Struct. Biol. 160 (2007) 35-48

We describe the development of quantitative electron spectroscopic tomography (QuEST), which provides 3-D distributions of elements on a nanometer scale. Specifically, it is shown that QuEST can be applied to map the distribution of phosphorus in unstained sections of embedded cells. A series of 2-D elemental maps is derived from images recorded in the energy filtering transmission electron microscope for a range of specimen tilt angles. A quantitative 3-D elemental distribution is then reconstructed from the elemental tilt series. To obtain accurate quantitative elemental distributions it is necessary to correct for plural inelastic scattering at the phosphorus L_2,3 edge, which is achieved by acquiring unfiltered and zero-loss images at each tilt angle. The data are acquired automatically using a cross correlation technique to correct for specimen drift and focus change between successive tilt angles. An algorithm based on the simultaneous iterative reconstruction technique (SIRT) is implemented to obtain quantitative information about the number of phosphorus atoms associated with each voxel in the reconstructed volume. We assess the accuracy of QuEST by determining the phosphorus content of ribosomes in a eukaryotic cell, and then apply it to estimate the density of nucleic acid in chromatin of the cell’s nucleus. From our experimental data, we estimate that the sensitivity for detecting phosphorus is 20 atoms in a 2.7 nm-sized voxel.     

The effect of attenuation map,scatter energy window width,and volume of interest on the calibration factor calculation in quantitative 177Lu SPECT imaging: Simulation and phantom study

PurposeThe objective of this study was to evaluate the image degrading factors in quantitative ¹⁷⁷Lu SPECT imaging when using both main gamma photopeak energies.MethodsPhantom measurements with two different vials containing various calibrated activities in air or water were performed to derive a mean calibration factor (CF) for large and small volumes of interest (VOIs). In addition, Monte Carlo simulations were utilized to investigate the effect of scatter energy window width, scatter correction method, such as effective scatter source estimation (ESSE) and triple energy window (TEW), and attenuation map on the quantification of ¹⁷⁷Lu. Results: The measured mean CF using large and small VOIs in water was 4.50 ± 0.80 and 4.80 ± 0.72 cps MBq⁻¹, respectively. Simulations showed a reference CF of 3.3 cps MBq⁻¹ for the water-filled phantom considering all photons excluding scattered events. By using the attenuation map generated for 190 keV photons, the calculated CFs for 113 keV and 208 keV are 10% lower than by using the weighted mean energy of 175 keV for ¹⁷⁷Lu. The calculated CF using the TEW correction was 17% higher than using the ESSE method for a water-filled phantom. However, our findings showed that an appropriate scatter window combination can reduce this difference between TEW and ESSE methods.ConclusionsThe present work implies that choosing a suitable width of scatter energy windows can reduce uncertainties in radioactivity quantification. It is suggested to generate the attenuation map at 113 keV and 208 keV, separately. Furthermore, using small VOIs is suggested in CF calculation.     

Using polarization analysis to separate the coherent and incoherent scattering from protein samples

Polarization analysis was used to separate experimentally the coherent and spin-incoherent nuclear static scattering functions, from a representative set of samples of interest for protein studies. This method had so far limited application in the study of amorphous materials, despite the relevance of the information that it provides. It allows, for instance, the experimental determination of the structure factor of materials containing a significant amount of hydrogen atoms, avoiding the contamination of measurements by a non-negligible incoherent background. Knowledge of the relative importance of the coherent and incoherent terms at different Q-values is also a pre-requisite for the interpretation of quasielastic neutron scattering experiments, performed at instruments in which the total dynamic scattering function is measured, such as conventional time-of-flight and backscattering spectrometers. Combining data from different instruments, it was possible to cover a wide Q-range, from the small-angle region (0.006 < Q < 0.04 Å^− 1) to the wide-angle region (up to ≈ 2.35 Å^− 1). Quantitative information was obtained on the fraction of coherent to spin-incoherent scattering from different protein samples: deuterated and protonated protein powders at different hydration levels and solutions of protonated proteins in D₂O at different concentrations. The results obtained are discussed in the context of the validity of the assumptions generally made when interpreting quasielastic neutron scattering experiments performed without polarization analysis.     

Cerenkov radiation energy transfer (CRET) imaging: a novel method for optical imaging of PET isotopes in biological systems

Background

Positron emission tomography (PET) allows sensitive, non-invasive analysis of the distribution of radiopharmaceutical tracers labeled with positron (β⁺)-emitting radionuclides in small animals and humans. Upon β⁺ decay, the initial velocity of high-energy β⁺ particles can momentarily exceed the speed of light in tissue, producing Cerenkov radiation that is detectable by optical imaging, but is highly absorbed in living organisms.

Principal Findings

To improve optical imaging of Cerenkov radiation in biological systems, we demonstrate that Cerenkov radiation from decay of the PET isotopes ⁶⁴Cu and ¹⁸F can be spectrally coupled by energy transfer to high Stokes-shift quantum nanoparticles (Qtracker705) to produce highly red-shifted photonic emissions. Efficient energy transfer was not detected with ^99mTc, a predominantly γ-emitting isotope. Similar to bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), herein we define the Cerenkov radiation energy transfer (CRET) ratio as the normalized quotient of light detected within a spectral window centered on the fluorophore emission divided by light detected within a spectral window of the Cerenkov radiation emission to quantify imaging signals. Optical images of solutions containing Qtracker705 nanoparticles and [¹⁸F]FDG showed CRET ratios in vitro as high as 8.8±1.1, while images of mice with subcutaneous pseudotumors impregnated with Qtracker705 following intravenous injection of [¹⁸F]FDG showed CRET ratios in vivo as high as 3.5±0.3.

Conclusions

Quantitative CRET imaging may afford a variety of novel optical imaging applications and activation strategies for PET radiopharmaceuticals and other isotopes in biomaterials, tissues and live animals.     

Influence du positionnement du patient sur la survenue des artefacts et l’interprétation des images de perfusion myocardique obtenues à partir d’une caméra dédiée à semi-conducteurs CZT

Myocardial perfusion imaging (MPI) can be affected by physical artefacts, such as attenuation and Compton scatter, both related to patient's morphological characteristics and his/her position during acquisition. These positional artefacts have been largely studied with classical Anger cameras. Recently, a new generation of heart-dedicated cameras has emerged, using novel cadmium–zinc–telluride (CZT) semi-conductors, with different physical characteristics and geometrical distribution. Our study aimed to investigate, in this new generation camera, the relation between patient's positioning and the occurring of physical artefacts. Plus, we examined whether these artefacts were likely to affect the interpretation made with such images.Material and methodsIn this prospective study, we included 60 patients scheduled in our unit for MPI. All of them underwent both prone and supine acquisitions, after stress test as well as at rest.ResultsAt a global population level, we observed significantly more abnormalities in supine acquisitions in territories mainly vascularized by right coronary artery, and to a lesser extent, by circumflex artery, than observed in prone acquisitions. No scoring difference was observed, regardless of position, in Summed Stress Score, Summed Rest Score or Summed Difference Score. This observation remained the same whether on a global population level, or on a BMI- and perimeter-basis level.ConclusionPosition related physical artefacts, such as attenuation and Compton scattering, are similar to those previously described with Anger cameras. These artefacts do not affect the interpretation of myocardial scans significantly, provided that both images are acquired in the same anatomical position.     

Animated visualization of a high resolution color three dimensional digital computer model of the whole human head

The interactive visualization of animated images through a computerized three dimensional (3D) full color model of an unstained cadaveric human head is presented. Serial full color images were taken of the blockface of a cryomicrotomed frozen human head every 200 μm. From this series of images a three dimensional digital model with a resultant pixel resolution of 200 μm³ was created on a UNIX workstation. Using this database, resampled images were computed along orthogonal axes and written sequentially to a write-once-read-many times (WORM) videodisc unit. Playback of this customized videodisc dataset provides animations of the digitally reconstructed slices and 3D reconstructed surface models. An interactive interface to the animated sequences is provided through a PC based tutorial package. This tutorial program is able to access videodisc frames to display animations and labeled still images in a software window to illustrate various neuroanatomic topics. The technique of animation as applied to this high resolution 3D model provides insight into complex spatial relationships and has great potential in research and as a teaching tool in the neurosciences.