A three-dimensional quality-guided phase unwrapping method for MR elastography

Magnetic resonance elastography (MRE) uses accumulated phases that are acquired at multiple, uniformly spaced relative phase offsets, to estimate harmonic motion information. Heavily wrapped phase occurs when the motion is large and unwrapping procedures are necessary to estimate the displacements required by MRE. Two unwrapping methods were developed and compared in this paper. The first method is a sequentially applied approach. The three-dimensional MRE phase image block for each slice was processed by two-dimensional unwrapping followed by a one-dimensional phase unwrapping approach along the phase-offset direction. This unwrapping approach generally works well for low noise data. However, there are still cases where the two-dimensional unwrapping method fails when noise is high. In this case, the baseline of the corrupted regions within an unwrapped image will not be consistent. Instead of separating the two-dimensional and one-dimensional unwrapping in a sequential approach, an interleaved three-dimensional quality-guided unwrapping method was developed to combine both the two-dimensional phase image continuity and one-dimensional harmonic motion information. The quality of one-dimensional harmonic motion unwrapping was used to guide the three-dimensional unwrapping procedures and it resulted in stronger guidance than in the sequential method. In this work, in vivo results generated by the two methods were compared.     

Towards optimized MR thermometry of the human heart at 3T

Catheter ablation using radio frequency (RF) has been used increasingly for the treatment of cardiac arrhythmias and may be combined with proton resonance frequency shift (PRFS) ?based MR thermometry to determine the therapy endpoint. We evaluated the suitability of two different MR thermometry sequences (TFE and TFE‐EPI) and three blood suppression techniques. Experiments were performed without heating, using an optimized imaging protocol including navigator respiratory compensation, cardiac triggering, and image processing for the compensation of motion and susceptibility artefacts. Blood suppression performance and its effect on temperature stability were evaluated in the ventricular septum of eight healthy volunteers using multislice double inversion recovery (MDIR), motion sensitized driven equilibrium (MSDE), and inflow saturation by saturation slabs (IS). It was shown that blood suppression during MR thermometry improves the contrast‐to‐noise ratio (CNR), the robustness of the applied motion correction algorithm as well as the temperature stability. A gradient echo sequence accelerated by an EPI readout and parallel imaging (SENSE) and using inflow saturation blood suppression was shown to achieve the best results. Temperature stabilities of 2 °C or better in the ventricular septum with a spatial resolution of 3.5 × 3.5 × 8mm³ and a temporal resolution corresponding to the heart rate of the volunteer, were observed. Our results indicate that blood suppression improves the temperature stability when performing cardiac MR thermometry. The proposed MR thermometry protocol, which optimizes temperature stability in the ventricular septum, represents a step towards PRFS‐based MR thermometry of the heart at 3 T. Copyright © 2011 John Wiley & Sons, Ltd.     

FID navigator‐based MR thermometry method to monitor small temperature changes in the brain of ventilated animals

An MR thermometry method is proposed for measuring in vivo small temperature changes engendered by external RF heat sources. The method relies on reproducible and stable respiration and therefore currently applies to ventilated animals whose breathing is carefully controlled. It first consists in characterizing the stability of the main magnetic field as well as the variations induced by breathing during a first monitoring stage. Second, RF heating is applied while the phase and thus temperature evolutions are continuously measured, the corrections due to breathing and field drift being made thanks to the data accumulated during the first period. The RF heat source is finally stopped and the temperature rise likewise is continuously monitored during a third and last stage to observe the animal cooling down and to validate the assumptions made for correcting for the main field variation and the physiological noise. Experiments were performed with a clinical 7 T scanner on an anesthetized baboon and with a dedicated RF heating setup. Analysis of the data reveals a precision around 0.1°C, which allows us to reliably measure sub‐degree temperature rises in the muscle and in the brain of the animal. Copyright © 2014 John Wiley & Sons, Ltd.     

A rapid immunostaining method utilizing preformed antibody-avidin-biotin-peroxidase complexes

A rapid immunostaining method utilizing a preformed antibody-avidin-biotin-peroxidase complex that produces staining in paraffin sections and in frozen sections of tissues in 15 minutes is described. Although the complex did not produce staining with the common leukocyte monoclonal antibody, consistent staining was seen with most antibodies used. The complex gave consistent and reproducible staining for at least three months when maintained in storage at 4 degrees C. The results show that many available antibodies can be adapted to a rapid one-step staining procedure employing the avidin-biotin-peroxidase complex system.     

A method for I131 conversion ratio utilizing the plasma inorganic I131 fraction

A method for calculating I¹³¹ conversion ratios by utilizing the inorganic I¹³¹ counts in the plasma at 24 hours is presented. Its chief advantage over previously used methods is that in cases of low conversion ratio it eliminates the large error introduced by fluctuations in the background of the counting equipment. The principle of the method depends upon the ability of silver to precipitate iodine (I¹³¹) in the presence of a slight excess of carrier iodine. Data are presented which show that low conversion ratios are frequently found in euthyroid patients, as has been the finding of other investigators.     

Joint edge detection and motion estimation of cardiac MR image sequence by a phase field method

In this paper a variational framework for joint segmentation and motion estimation is employed for inspecting heart in Cine MRI sequences. A functional including Mumford-Shah segmentation and optical flow based dense motion estimation is approximated using the phase-field technique. The minimizer of the functional provides an optimum motion field and edge set by considering both spatial and temporal discontinuities. Exploiting calculus of variation principles, multiple partial differential equations associated with the Euler-Lagrange equations of the functional are extracted, first. Next, the finite element method is used to discretize the resulting PDEs for numerical solution. Several simulation runs are used to test the convergence and the parameter sensitivity of the method. It is further applied to a comprehensive set of clinical data in order to compare with conventional cascade methods. Developmental constraints are identified as memory usage and computational complexities, which may be resolved utilizing sparse matrix manipulations and similar techniques. Based on the results of this study, joint segmentation and motion estimation outperforms previously reported cascade approaches especially in segmentation. Experimental results substantiated that the proposed method extracts the motion field and the edge set more precisely in comparison with conventional cascade approaches. This superior result is the consequence of simultaneously considering the discontinuity in both motion field and image space and including consequent frames (usually five) in our joint process functional.     

A Nycodenz gradient method for the purification of neutrophils from the peripheral blood of rats

A Nycodenz gradient technique is described which permits the separation of functionally active polymorphonuclear neutrophils (PMN) from the peripheral blood of rats. PMN are obtained at greater than 90% purity and fractionate at a peak density of 1.0919 g/ml. The method is suitable for isolating PMN when the circulating PMN count is low (less than 20%) as in normal rats and when the count is high (greater than 30%), as a result of inducing inflammation in the subcutaneous air pouch of rats by the intra-pouch injection of peptone. The chemotactic responsiveness of rat PMN was found to be markedly less than that of human PMN when the formyl peptides were used as chemoattractants but not when zymosan-activated serum was the chemoattractant. Blood PMN from normal rats and from peptone-treated rats showed no significant difference in their response to ZAS indicating that priming of their activity by the induction of long term (8 day) inflammation was not a feature of these experiments. However, air pouch-derived PMN displayed a highly significant reduction in activity compared with their isologous blood-PMN. The Nycodenz method offers an alternative to the Percoll separation method for rat blood and will be useful when comparative studies of elicited and non-elicited PMN are required since the latter are obtained in sufficiently high yield and purity for microassays on their function to be performed.     

An improved method for the quantitation of A2 hemoglobin utilizing cellulose acetate electrophoresis and densitometry.

An improved method of A2 quantitation by cellulose acetate electrophoresis is describes. Densitometry is performed on uncleared membranes. The height of the peak obtained for the A2 band is compared with a standard curve derived from peaks obtained by serial dilutions of hemoglobin. Noral A2 hemoglobin as measured by this method is 2.55 percent plus or minus 0.60 (2 S.D.). Twenty patients samples were run in parallel with a column chromatographic method. Eight samples were found to contain elevated A2 levels by the column method. The same eight samples were correctly identified by the electrophoretic technic.     

A new method for separating lymphocytes and granulocytes from human peripheral blood using programmed gradient sedimentation in an isokinetic gradient

Isopycnic sedimentation and programmed rate-zonal sedimentation in an isokinetic gradient of Ficoll (polysucrose) in tissue culture medium were used for the purification of individual kinds of cells from human peripheral blood. Following centrifugation in the isokinetic gradient, lymphocytes were 99.9±0.1 per cent and granulocytes 90.1±3.8 per cent of nucleated cells in their respective fractions. While monocytes were concentrated in one zone of the gradient, they were not as highly purified as monocytes prepared using established methods. Purification of lymphocytes and granulocytes using the isokinetic gradient offers one of the most effective and rapid means for obtaining purified sterile lymphocytes and granulocytes for in vitro or biochemical studies.     

A photoelectric diameter gauge utilizing the image sensor

We have designed and constructed a blood vessel diameter gauge, utilizing a high resolving power and stability of the image sensor which has become of major interest lately as a new photoelectric transducer. The distinct feature of the gauge is summarized as follows. (1) Contact-free measurements of blood vessel diameter (1–14 mm) are possible. (2) The cuff-type probe is small in size and easy to put to use. (3) The lightguide consisting of optical fibers makes incident light cold. (4) The resolving power of the gauge is 28 m in principle. (5) The frequency response of the gauge is flat up to 100 Hz. A few results of its preliminary application to in vitro and in vivo experiments are also demonstrated.     

Application of the cell block method, utilizing mount quick mounting medium

Our objective was to determine the applicability of cell transfer and cell block methods using Mount Quick (Daido Sangyo, Saitama, Japan) mounting medium (MQ) for hematoxylin-eosin (H&E) and immunohistochemical staining of several limited amounts of biological materials in slide preparations. The materials investigated were histopathologically confirmed malignant mesotheliomas (pleural effusions) and malignant lymphomas, a malignant melanoma, and an amelanotic melanoma in sealed slides. Monoclonal antibodies against carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), cancer antigen 125 (CA-125), vimentin, thrombomodulin (TM), cytokeratin, UCHL-1, L-26, melanoma-specific antigen (HMB45), and S-100 protein (S-100) were applied in the investigation. The malignant mesotheliomas were found to be positive for EMA, cytokeratin, vimentin, TM, and CA-125, and negative for CEA, with no differences being observed in findings from direct contact preparations. Using T-cell-type malignant lymphomas for immunohistochemistry, UCHL-1 positivity and L-26 negativity were clearly demonstrated. The malignant melanoma and amelanotic melanoma materials stained strongly for HMB45 and S-100. Cell transfer employing MQ is a suitable approach for immunohistochemical investigations of limited materials. In addition, cell blocks derived from MQ-embedded smears can be used for both H&E and immunohistochemical staining. Diagn. Cytopathol. 2000;22:117-119.     

A novel method for accurate collagen and biochemical assessment of pulmonary tissue utilizing one animal

Aim: The purpose of this study was to develop an improved method for collagen and protein assessment of fibrotic lungs while decreasing animal use. methods: 8-10 week old, male C57BL/6 mice were given a single intratracheal instillation of crocidolite asbestos or control titanium dioxide. Lungs were collected on day 14 and dried as whole lung, or homogenized in CHAPS buffer, for hydroxyproline analysis. Insoluble and salt-soluble collagen content was also determined in lung homogenates using a modified Sirius red colorimetric 96-well plate assay. results: The hydroxyproline assay showed significant increases in collagen content in the lungs of asbestos-treated mice. Identical results were present between collagen content determined on dried whole lung or whole lung homogenates. The Sirius red plate assay showed a significant increase in collagen content in lung homogenates however, this assay grossly over-estimated the total amount of collagen and underestimated changes between control and fibrotic lungs, conclusions: The proposed method provides accurate quantification of collagen content in whole lungs and additional homogenate samples for biochemical analysis from a single animal. The Sirius-red colorimetric plate assay provides a complementary method for determination of the relative changes in lung collagen but the values tend to overestimate absolute values obtained by the gold standard hydroxyproline assay and underestimate the overall fibrotic injury.     

Evaluation of an amylase method utilizing p-nitrophenyl glucosides as substrates

An enzymatic method which uses p-nitrophenyl glucosides as substrates for the determination of amylase activity has been evaluated. The kit can be used in either a two point or multiple point mode but these give different results. In both modes the reagent gave a linear response to at least 800 U/I. No interference by endogenous compounds was observed as has been reported with some enzymatic procedures. Within-batch precision (coefficient of variation 1% at 340 U/I and 5% at 10 U/I) and inter-batch precision (coefficient of variation 3.4% at 170 U/I and 3.7% at 56 U/I) are comparable to other amylase methodologies. Correlation with both the manual dye-starch (Phadebas) (r2 = 0.994) and the Beckman Amylase-DS (r2 = 0.998) methods was good for the sera and urines examined. Normal range for serum was 17 to 84 U/I (mean +/- 2 SD). The method is rapid (assay time 10 min), requires very little specimen (20 microliters for the two point assay and 5 microliters for the multiple point assay) and is suitable for both routine and emergency use.     

A rapid method to isolate platelets from human blood by density gradient centrifugation.

Platelets can be damaged easily or activated during isolation, making them unsuitable for functional studies. The most common technique for isolating platelets involves centrifugation. Although gentler methods have been devised to isolate platelets by density gradient centrifugation or electrophoresis, these techniques either result in a relatively dilute platelet preparation or are time-consuming. A simple, gentle technique for isolating concentrated platelet preparations for experimental or clinical use is reported. Freshly drawn whole blood was spun over a commercially available density gradient medium for 30 minutes. The mononuclear cell layer (which also contains most of the platelets) was collected and nucleated cells were pelleted by centrifugation. The recovery of platelets was about 60%. Contamination with leukocytes was less than 1%, and the platelet concentration was about 130% of blood concentration. Higher concentrations can be obtained if more whole blood is layered onto the Mono-Poly Resolving Medium (MPRM; Flow Laboratories, McLean, VA). About 10% of the platelets expressed the activation marker GMP-140 by flow cytometric analysis. They could be activated by thrombin so that 70% to 90% of the platelets expressed GMP-140. Thus, this technique can rapidly and easily yield a functionally intact platelet preparation. This preparation can be purified again if needed. No specialized skills or equipment are needed. A significant advantage of the method is that platelets can be obtained from thrombocytopenic patients in final concentrations that are high enough to use for platelet function testing.