Commissioning of a solid tank design for fan-beam optical CT based 3D radiation dosimetry.

Objective.Optical computed tomography (CT) is one of the leading modalities for imaging gel dosimeters used in the verification of complex radiotherapy treatments. In previous work, a novel fan-beam optical CT scanner design was proposed that could significantly reduce the volume of the refractive index baths that are commonly found in optical CT systems. Here, the proposed scanner has been manufactured and commissioned.Approach.Image reconstruction is performed through algebraic reconstruction technique and iterated using the fast iterative shrinkage-thresholding algorithm (FISTA) algorithm. Ray tracing for algebraic reconstruction was performed using an in-house developed ray tracing simulator. A set of Sylgard® 184 phantoms were created to commission spatial resolution, geometric deformity, contrast-to-noise ratio (CNR), and scan settings.Main Results.The scanner is capable of a 0.929 mm-1spatial resolution, observed at 200 iterations, although the spatial resolution is highly dependent on the number of iterations. The geometric distortion, measured by scanning a needle phantom with the prototype scanner as well as a conventional x-ray CT was found to be within <0.25 mm. The CNR was found to peak between 65 and 190 occurring between 50 and 100 iterations and was highly dependent on the region chosen for background noise calculation. The proposed scanner is capable of scanning and reading out slices in less than 1 min per slice.Significance.This work displays the viability of a fan-beam optical CT scanner with minimal index matching using ray-traced algebraic reconstruction.

Evaluation of an x-ray CT polymer gel dosimetry system in the measurement of deformed dose.

This study is an evaluation of the use of a N-isopropylacrylamide (NIPAM)-based x-ray CT polymer gel dosimetry (PGD) system in the measurement of deformed dose. This work also compares dose that is measured by the gel dosimetry system to dose calculated by a novel deformable dose accumulation algorithm, defDOSXYZnrc, that uses direct voxel tracking. Deformable gels were first irradiated using a single 3.5 × 5 cm2 open field and the static dose was compared to defDOSXYZnrc as a control measurement. Gel measurement was found to be in excellent agreement with defDOSXYZnrc in the static case with gamma passing rates of 94.5% using a 3%/3 mm criterion and 93.3% using a 3%/2 mm criterion. Following the static measurements, a deformable gel was irradiated with the same single field under an external compression of 25 mm and then released from this compression for dosimetric read out. The measured deformed dose was then compared to deformed dose calculated by defDOSXYZnrc based on deformation vectors produced by the Velocity AI deformable image registration (DIR) algorithm. In the deformed dose distribution there were differences in the measured and calculated field position of up to 0.8 mm and differences in the measured in calculated field size of up to 11.9 mm. Gamma pass rates were 60.0% using a 3%/3 mm criterion and 56.8% using a 3%/2 mm criterion for the deforming measurements representing a decrease in agreement compared to the control measurements. Further analysis showed that passing rates increased to 86.5% using a 3%/3 mm criterion and 70.5% using a 3%/2 mm criterion in voxels within 5 mm of fiducial markers used to guide the deformable image registration. This work represents the first measurement of deformed dose using x-ray CT polymer gel dosimetry. Overall these results highlight some of the challenges in the calculation and measurement of deforming dose and provide insight into possible strategies for improvement.

Linac-integrated kV-cone beam CT polymer gel dosimetry.

X-ray CT polymer gel dosimetry (PGD) remains a promising tool for three dimensional verification of high-dose treatment deliveries such as non-coplanar stereotactic irradiations. Recent demonstrations have shown a proof-of-principle application of linac-integrated cone beam CT-imaged (LI-CBCT) PGDs for 3D dose verification. LI-CBCT offers advantages over previous CT based PGD, including close to real-time imaging of the irradiated dosimeter, as well as the ability to maintain the dosimeter in the same physical location for irradiation and imaging, thereby eliminating spatial errors due to dosimeter re-positioning for read-out that may occur for other systems. However the dosimetric characteristics of a LI-CBCT PGD system remain to be established. The work herein determines the dosimetric properties and critical parameters needed to perform cone beam PGD. In particular, we show that imaging the dosimeter 20-30 min post irradiation offers excellent recovery of maximum polymerization yield ([Formula: see text]90%), averaging with as few as 10 image averages can provide ∼90% gamma pass rates (3%, 3 mm) as compared to treatment planning, and that eliminating outlier averaging points can improve the precision and signal to noise ratio of resultant images. In summary, with appropriate methodology LI-CBCT PGD can provide dosimetric data capable of verification of complex high dose radiation deliveries in three dimensions and may find use in commissioning and validation of novel complex treatments.

Optimization of solid tank design for fan-beam optical CT based 3D radiation dosimetry.

Optical computed tomography (CT) is one of the leading modalities for imaging gel dosimeters for 3D radiation dosimetry. There exist multiple scanner designs that have showcased excellent 3D dose verification capabilities of optical CT gel dosimetry. However, due to multiple experimental and reconstruction based factors there is currently no single scanner that has become a preferred standard. A significant challenge with setup and maintenance can be attributed to maintaining a large refractive index bath (1-15 l). In this work, a prototype solid 'tank' optical CT scanner is proposed that minimizes the volume of refractive index bath to between 10 and 35 ml. A ray-path simulator was created to optimize the design such that the solid tank geometry maximizes light collection across the detector array, maximizes the volume of the dosimeter scanned, and maximizes the collected signal dynamic range. An objective function was created to score possible geometries, and was optimized to find a local maximum geometry score from a set of possible design parameters. The design parameters optimized include the block length x bl , bore position x bc , fan-laser position x lp , lens block face semi-major axis length x ma , and the lens block face eccentricity x be . For the proposed design it was found that each of these parameters can have a significant effect on the signal collection efficacy within the scanner. Simulations scores are specific to the attenuation characteristics and refractive index of a simulated dosimeter. It was found that for a FlexyDos3D dosimeter, the ideal values for each of the five variables were: x bl = 314 mm, x bc = 6.5 mm, x lp = 50 mm, x ma = 66 mm, and x be = 0. In addition, a ClearView™ dosimeter was found to have ideal values at: x bl = 204 mm, x bc = 13 mm, x lp = 58 mm, x ma = 69 mm, and x be = 0. The ray simulator can also be used for further design and testing of new, unique and purpose-built optical CT geometries.

Effects of glycerol co-solvent on the rate and form of polymer gel dose response.

A factor currently limiting the clinical utility of x-ray CT polymer gel dosimetry is the overall low dose sensitivity (and hence low dose resolution) of the system. Hence, active research remains in the investigation of polymer gel formulations with increased CT dose response. An ideal polymer gel dosimeter will exhibit a sensitive CT response which is linear over a suitable dose range, making clinical implementation reasonably straightforward. This study reports on the variations in rate and form of the CT dose response of irradiated polymer gels manufactured with glycerol, which is a co-solvent that permits dissolution of additional bisacrylamide above its water solubility limit (3% by weight). This study focuses on situations where the concentration of bisacrylamide is kept at or below its water solubility limit so that the influence of the co-solvent on the dose response can be explored separately from the effects of increased cross-linker concentration. CT imaging and Raman spectroscopy are used to construct dose-response curves for irradiated gels varying in (i) initial total monomer (%T) and (ii) initial co-solvent concentration. Results indicate that: (i) for a fixed glycerol concentration, gel response increases linearly with %T. Furthermore, the functional form of the dose response remains constant, in agreement with a previous model of polymer formation. (ii) Polymer gels with constant %T and increasing co-solvent concentrations also show enhanced CT response. In addition, the functional form of the response is altered in these gels as co-solvent concentration is increased. Raman data indicate that the fraction of bis-acrylamide incorporated into polymerization, as opposed to cyclization, increases as co-solvent concentration increases. The changes in functional form indicate varying polymer yields (per unit dose), akin to relative fractional monomer/cross-linker (i.e. %C) changes in earlier studies. These results are put into context of the model of polymer formation. The implications of these results on the clinical utility of polymer gels with co-solvent are highlighted.

Preliminary investigation of the NMR, optical and x-ray CT dose-response of polymer gel dosimeters incorporating cosolvents to improve dose sensitivity.

This study reports on efforts to increase the dose sensitivity of polymer gel dosimeters used in 3D radiation dosimetry. The potential of several different cosolvents is investigated, with the aim of increasing the solubility of N,N'-methylene-bisacrylamide crosslinker in polymer gel dosimeters. Glycerol and isopropanol increase the limit for the crosslinker solubility from approximately 3% to 5% and 10% by weight, respectively. This enables the manufacture of polymer gel dosimeters with much higher levels of crosslinking than was previously possible. New dosimeter recipes containing up to 5 wt% N,N'-methylene-bisacrylamide were subjected to spatially uniform radiation and were studied using nuclear magnetic resonance (NMR), as well as x-ray and optical CT techniques. The resulting dosimeters exhibit dose sensitivities that are up to 2.7 times higher than measured for a typical dosimeters with 3% N,N'-methylene-bisacrylamide without the addition of cosolvent. Two additional cosolvents (n-propanol and sec-butanol) were deemed unsuitable for practical dosimeters due to incompatibility with gelatin, cloudiness prior to irradiation, and immiscibility with water when large quantities of cosolvent were used. The dosimeters with high N,N'-methylene-bisacrylamide content that used isopropanol or glycerol as cosolvents had high optical clarity prior to irradiation, but did not produce suitable optical CT results for non-uniformly irradiated gels due to polymer development outside of the high dose regions of the pencil beams and significant light scatter. Further experiments are required to determine whether cosolvents can be used to manufacture gels with sufficiently high dose sensitivity for readout using x-ray computed tomography.

The use of ultraviolet resonance Raman spectroscopy in the analysis of ionizing-radiation-induced damage in DNA.

Ultraviolet resonance Raman spectroscopy (UVRRS) was used to determine damage done in both calf-thymus DNA (CT-DNA) and a short stranded DNA oligomer (SS-DNA) due to ionizing radiation from a medical (60)Co radiation therapy unit used in the treatment of cancer. Spectra were acquired at incident UV wavelengths of 248, 257, and 264 nm in order to utilize the differences in UVRR cross-sections of the bases with wavelength. Through the analysis of difference spectra between irradiated and unirradiated DNA at each of the incident UV wavelengths, damage to CT- and SS-DNA was observed and identified. Significant radiation-induced increases in the difference spectra of the CT-DNA indicated disruption of the stable, stacked structure of its bases, as well as the disruption of Watson-Crick hydrogen bonds between the base pairs. Base unstacking was not as evident in the SS-DNA, while radiation-induced spectral decreases suggest disruption of the structure of the nucleotides. As demonstrated, UVRRS has the ability to highlight contributions from specific moieties with the use of varying incident UV wavelengths, thus enhancing the already information-rich content of the Raman spectra.

Introduction of a deformable x-ray CT polymer gel dosimetry system.

This study introduces the first 3D deformable dosimetry system based on x-ray computed tomography (CT) polymer gel dosimetry and establishes the setup reproducibility, deformation characteristics and dose response of the system. A N-isopropylacrylamide (NIPAM)-based gel formulation optimized for x-ray CT gel dosimetry was used, with a latex balloon serving as the deformable container and low-density polyethylene and polyvinyl alcohol providing additional oxygen barrier. Deformable gels were irradiated with a 6 MV calibration pattern to determine dosimetric response and a dosimetrically uniform plan to determine the spatial uniformity of the response. Wax beads were added to each gel as fiducial markers to track the deformation and setup of the gel dosimeters. From positions of the beads on CT images the setup reproducibility and the limits and reproducibility of gel deformation were determined. Comparison of gel measurements with Monte Carlo dose calculations found excellent dosimetric accuracy, comparable to that of an established non-deformable dosimetry system, with a mean dose discrepancy of 1.5% in the low-dose gradient region and a gamma pass rate of 97.9% using a 3%/3 mm criterion. The deformable dosimeter also showed good overall spatial dose uniformity throughout the dosimeter with some discrepancies within 20 mm of the edge of the container. Tracking of the beads within the dosimeter found that sub-millimetre setup accuracy is achievable with this system. The dosimeter was able to deform and relax when externally compressed by up to 30 mm without sustaining any permanent damage. Internal deformations in 3D produced average marker movements of up to 12 mm along the direction of compression. These deformations were also shown to be reproducible over 100 consecutive deformations. This work has established several important characteristics of a new deformable dosimetry system which shows promise for future clinical applications, including the validation of deformable dose accumulation algorithms.

X-ray CT dose in normoxic polyacrylamide gel dosimetry.

This study reports on the effects of x-ray CT dose in CT imaged normoxic polyacrylamide (nPAG) gel dosimeters. The investigation is partitioned into three sections. First, the CT dose absorbed in nPAG is quantified under a range of typical gel CT imaging protocols. It is found that the maximum absorbed CT dose occurs for volumetric imaging and is in the range of 4.6 +/- 0.2 cGy/image. This does scales linearly with image averaging. Second, using Raman spectroscopy, the response of nPAG to CT imaging photon energies (i.e., 120-140 kVp) is established and compared to the well known dose response of nPAG exposed to 6 MV photons. It is found that nPAG exhibits a weaker response (per unit dose) to 140-kVp incident photons as compared to 6 MV incident photons (slopes m6 mv = -0.0374 +/- 0.0006 Gy(-1) and m140 kVp = -0.016 +/- 0.001 Gy(-1)). Finally, using the above data, an induced change in CT number (deltaN(CT)) is calculated for nPAG imaged using a range of gel imaging protocols. It is found that under typical imaging protocols (120-140 kVp, 200 mAs, approximately 16-32 image averages) a deltaN(CT) < 0.2 H is induced in active nPAG dosimeters. This deltaN(CT) is below the current limit of detectability of CT nPAG polymer gel dosimetry. Under expanded imaging protocols (e.g., very high number of image averages) an induced deltaN(CT) of approximately 0.5 H is possible. In these situations the additional polymerization occurring in nPAG due to the imaging process may need to be accounted for.

Improving the quality of reconstructed X-ray CT images of polymer gel dosimeters: zero-scan coupled with adaptive mean filtering.

This study evaluated the feasibility of combining the 'zero-scan' (ZS) X-ray computed tomography (CT) based polymer gel dosimeter (PGD) readout with adaptive mean (AM) filtering for improving the signal to noise ratio (SNR), and to compare these results with available average scan (AS) X-ray CT readout techniques. NIPAM PGD were manufactured, irradiated with 6 MV photons, CT imaged and processed in Matlab. AM filter for two iterations, with 3 × 3 and 5 × 5 pixels (kernel size), was used in two scenarios (a) the CT images were subjected to AM filtering (pre-processing) and these were further employed to generate AS and ZS gel images, and (b) the AS and ZS images were first reconstructed from the CT images and then AM filtering was carried out (post-processing). SNR was computed in an ROI of 30 × 30 for different pre and post processing cases. Results showed that the ZS technique combined with AM filtering resulted in improved SNR. Using the previously-recommended 25 images for reconstruction the ZS pre-processed protocol can give an increase of 44% and 80% in SNR for 3 × 3 and 5 × 5 kernel sizes respectively. However, post processing using both techniques and filter sizes introduced blur and a reduction in the spatial resolution. Based on this work, it is possible to recommend that the ZS method may be combined with pre-processed AM filtering using appropriate kernel size, to produce a large increase in the SNR of the reconstructed PGD images.

Plasmonic labeling of subcellular compartments in cancer cells: multiplexing with fine-tuned gold and silver nanoshells.

Fine-tuned gold and silver nanoshells were produced via an entirely reformulated synthesis. The new method yielded ultramonodisperse samples, with polydispersity indexes (PI) as low as 0.02 and narrow extinction bands suited for multiplex analysis. A library of nanoshell samples with localized surface plasmon resonances (LSPR) spanning across the visible range was synthesized. Hyperspectral analysis revealed that the average scattering spectrum of 100 nanoshells matched closely to the spectrum of a single nanoshell, indicating an unprecedented low level of nanoparticle-to-nanoparticle variation for this type of system. A cell labeling experiment, targeting different subcellular compartments in MCF-7 human breast cancer cells, demonstrated that these monodisperse nanoparticles can be used as a multiplex platform for single cell analysis at the intracellular and extracellular level. Antibody-coated gold nanoshells targeted the plasma membrane, while silver nanoshells coated with a nuclear localization signal (NLS) targeted the nuclear membrane. A fluorescence counterstaining experiment, as well as single cell hyperspectral microscopy showed the excellent selectivity and specificity of each type of nanoparticle for its designed subcellular compartment. A time-lapse photodegradation experiment confirmed the enhanced stability of the nanoshells over fluorescent labeling and their capabilities for long-term live cell imaging.

Investigation of tetrakis hydroxymethyl phosphonium chloride as an antioxidant for use in x-ray computed tomography polyacrylamide gel dosimetry.

Of the antioxidants used to scavenge oxygen in polymer gel dosimeters, tetrakis (hydroxymethyl) phosphonium chloride (THPC) has been shown to hold great promise due to its rapid oxygen scavenging abilities. In this study we (a) investigate the use of THPC as an antioxidant for polyacrylamide gel (PAGAT) dosimeters used in conjunction with x-ray computed tomography (CT) and (b) work to establish the reaction mechanisms of THPC with the polymer gel constituents. We establish the dose response reproducibility of PAGAT dosimeters when imaged with CT and show that PAGAT dosimeters exhibit highly reproducible dose responses for a range of irradiation times post gel manufacture (2-6 h) and CT imaging times post gel irradiation (1-5 days). The THPC concentration within the gel leading to a maximized dose response and minimized O(2) inhibition of polymerization is found to be approximately 4.5 mM. We further assess the stability of PAGAT dosimeters by investigating the reactions of THPC with the individual gel constituents. The importance of utilizing deionized water in polymer gel manufacture is noted. We show that, while THPC remains unreactive with acrylamide and bis-acrylamide under unirradiated conditions, THPC can react with gelatin to increase the cross-linking of the gelatin matrix in unirradiated dosimeters. THPC reactions with gelatin can lead to the lower observed dose sensitivity of PAGAT (approximately 0.36 +/- 0.04 H Gy(-1)) as compared to polyacrylamide gels manufactured under anoxic conditions (approximately 0.83 +/- 0.03 H Gy(-1)). The reactions of THPC which lead to O(2) scavenging, and potential reactions of THPC with other gel constituents, are proposed.

Investigation of a 2D two-point maximum entropy regularization method for signal-to-noise ratio enhancement: application to CT polymer gel dosimetry.

This study presents a new method of image signal-to-noise ratio (SNR) enhancement by utilizing a newly developed 2D two-point maximum entropy regularization method (TPMEM). When utilized as an image filter, it is shown that 2D TPMEM offers unsurpassed flexibility in its ability to balance the complementary requirements of image smoothness and fidelity. The technique is evaluated for use in the enhancement of x-ray computed tomography (CT) images of irradiated polymer gels used in radiation dosimetry. We utilize a range of statistical parameters (e.g. root-mean square error, correlation coefficient, error histograms, Fourier data) to characterize the performance of TPMEM applied to a series of synthetic images of varying initial SNR. These images are designed to mimic a range of dose intensity patterns that would occur in x-ray CT polymer gel radiation dosimetry. Analysis is extended to a CT image of a polymer gel dosimeter irradiated with a stereotactic radiation therapy dose distribution. Results indicate that TPMEM performs strikingly well on radiation dosimetry data, significantly enhancing the SNR of noise-corrupted images (SNR enhancement factors >15 are possible) while minimally distorting the original image detail (as shown by the error histograms and Fourier data). It is also noted that application of this new TPMEM filter is not restricted exclusively to x-ray CT polymer gel dosimetry image data but can in future be extended to a wide range of radiation dosimetry data.

Design and application of 3D-printed stepless beam modulators in proton therapy.

A new method for the design of stepless beam modulators for proton therapy is described and verified. Simulations of the classic designs are compared against the stepless method for various modulation widths which are clinically applicable in proton eye therapy. Three modulator wheels were printed using a Stratasys Objet30 3D printer. The resulting depth dose distributions showed improved uniformity over the classic stepped designs. Simulated results imply a possible improvement in distal penumbra width; however, more accurate measurements are needed to fully verify this effect. Lastly, simulations were done to model bio-equivalence to Co-60 cell kill. A wheel was successfully designed to flatten this metric.

Technical considerations for implementation of x-ray CT polymer gel dosimetry.

Gel dosimetry is the most promising 3D dosimetry technique in current radiation therapy practice. X-ray CT has been shown to be a feasible method of reading out polymer gel dosimeters and, with the high accessibility of CT scanners to cancer hospitals, presents an exciting possibility for clinical implementation of gel dosimetry. In this study we report on technical considerations for implementation of x-ray CT polymer gel dosimetry. Specifically phantom design, CT imaging methods, imaging time requirements and gel dose response are investigated. Where possible, recommendations are made for optimizing parameters to enhance system performance. The dose resolution achievable with an optimized system is calculated given voxel size and imaging time constraints. Results are compared with MRI and optical CT polymer gel dosimetry results available in the literature.

Dose rate properties of NIPAM-based x-ray CT polymer gel dosimeters.

In this work we investigate radiation dose rate dependencies of N-isopropylacrylamide (NIPAM) based polymer gel dosimeters (PGDs) used in conjunction with x-ray computed tomography imaging for radiotherapy dose verification. We define four primary forms of dose rate variation: constant mean dose rate where beam on and beam off times both vary, variable mean dose rate where beam on time varies, variable mean dose rate where beam off time varies and machine dose rate (MU min(-1)). We utilize both small (20 mL) vials and large volume (1L) gel containers to identify and characterize dose rate dependence in NIPAM PGDs. Results indicate that all investigated constant and variable mean dose rates had negligible affect on PGD dose response with the exception of machine dose rates (100-600 MU min(-1)) which produced variations in dose response significantly lower than previously reported. Explanations of the reduced variability in dose response are given. It is also shown that NIPAM PGD dose response is not affected by variations in dose rate that may occur in modulated treatment deliveries. Finally, compositional changes in NIPAM PGDs are investigated as potential mitigating strategies for dose rate-dependent response variability.

Incorporating multislice imaging into x-ray CT polymer gel dosimetry.

PURPOSE: To evaluate multislice computed tomography (CT) scanning for fast and reliable readout of radiation therapy (RT) dose distributions using CT polymer gel dosimetry (PGD) and to establish a baseline assessment of image noise and uniformity in an unirradiated gel dosimeter. METHODS: A 16-slice CT scanner was used to acquire images through a 1 L cylinder filled with water. Additional images were collected using a single slice machine. The variability in CT number (NCT) associated with the anode heel effect was evaluated and used to define a new slice-by-slice background subtraction artifact removal technique for CT PGD. Image quality was assessed for the multislice system by evaluating image noise and uniformity. The agreement in NCT for slices acquired simultaneously using the multislice detector array was also examined. Further study was performed to assess the effects of increasing x-ray tube load on the constancy of measured NCT and overall scan time. In all cases, results were compared to the single slice machine. Finally, images were collected throughout the volume of an unirradiated gel dosimeter to quantify image noise and uniformity before radiation is delivered. RESULTS: Slice-by-slice background subtraction effectively removes the variability in NCT observed across images acquired simultaneously using the multislice scanner and is the recommended background subtraction method when using a multislice CT system. Image noise was higher for the multislice system compared to the single slice scanner, but overall image quality was comparable between the two systems. Further study showed NCT was consistent across image slices acquired simultaneously using the multislice detector array for each detector configuration of the slice thicknesses examined. In addition, the multislice system was found to eliminate variations in NCT due to increasing x-ray tube load and reduce scanning time by a factor of 4 when compared to imaging a large volume using a single slice scanner. Images acquired through an unirradiated, active gel revealed NCT varies between the top and bottom of the 1 L cylinder as well as across the diameter of the cylinder by up to 7 HU. CONCLUSIONS: Multislice CT imaging has been evaluated for CT PGD and found to be the superior technique compared to single slice imaging in terms of the time required to complete a scan and the tube load characteristics associated with each scanning method. The implementation of multislice scanning is straightforward and expected to facilitate routine gel dosimetry measurements for complex dose distributions in modern RT centers.

3D printed plastics for beam modulation in proton therapy.

Two 3D printing methods, fused filament fabrication (FFF) and PolyJet™ (PJ) were investigated for suitability in clinical proton therapy (PT) energy modulation. Measurements of printing precision, printed density and mean stopping power are presented. FFF is found to be accurate to 0.1 mm, to contain a void fraction of 13% due to air pockets and to have a mean stopping power dependent on geometry. PJ was found to print accurate to 0.05 mm, with a material density and mean stopping power consistent with solid poly(methyl methacrylate) (PMMA). Both FFF and PJ were found to print significant, sporadic defects associated with sharp edges on the order of 0.2 mm. Site standard PT modulator wheels were printed using both methods. Measured depth-dose profiles with a 74 MeV beam show poor agreement between PMMA and printed FFF wheels. PJ printed wheel depth-dose agreed with PMMA within 1% of treatment dose except for a distal falloff discrepancy of 0.5 mm.

Aleukemic granulocytic sarcoma with AML1/ETO fusion gene expression and clonal T cell populations.

We report a unique case of aleukemic granulocytic sarcoma of the neck, originally misdiagnosed as non-Hodgkin's lymphoma (NHL), though chloroma was also suspected due to a greenish macroscopic appearance and the presence of myeloid chloroacetate esterase (CAE)+ cells. The proof of clonal T cell receptor gamma chain (TcRgamma) gene rearrangements in the recurring tumor was deemed to confirm the initial diagnosis of T cell NHL. Altogether five distinct types of clonal TcRgamma gene rearrangements were found in the tumor, bone marrow and peripheral blood. Only retrospectively, using RT-PCR, did we detect the acute myeloid leukemia subset-specific fusion gene AML1/ETO in the frozen samples of the relapsed tumor, as well as in the otherwise unaffected bone marrow and peripheral blood (representing 'minimal initial disease' in the latter two samples). Simultaneous staining verified that the neoplastic CAE+ cells and CD45RO+ T cells were different populations.

Near-tetraploid poorly differentiated acute myeloid leukemia M0 diagnosed by short-term cultures with a phorbol ester TPA.

Leukemic blasts of two patients with acute leukemia exhibited similar characteristics. They were heterogeneous in size with a diameter of 14-30 microns in smears and unclassifiable by morphological, cytochemical, immunophenotypic and ultrastructural examinations. Cytogenetic examinations of both revealed a near-tetraploid karyotype. Blasts from both patients differentiated into macrophages in cultures with 10 ng/ml 12-O-tetradecanoylphorbol-13-acetate (TPA) which is a feature specific for myeloid blasts and the cases were thus classified as poorly differentiated acute myeloid leukemias (AML M0). Near-tetraploid poorly differentiated acute myeloid leukemias M0 seem to be a special category of AML in the morphologic, immunologic and cytogenetic (MIC) classification. The presence of very large blasts in the heterogeneous blast population in acute unclassified leukemias could be a morphological sign of near-tetraploid leukemias AML M0.