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PubMed1992

Alignment, classification, and three-dimensional reconstruction of single particles embedded in ice.

Cryo-electron microscopy of single biological particles poses new challenges to digital image processing due to the low signal-to-noise ratio of the data. New tools have been devised to deal with important aspects of 3-D reconstruction following the random-conical data collection scheme: (a) a new shift-invariant function has been derived, which promises to facilitate alignment and classification of single particle projections; (b) a new method of orientation search is proposed, which makes it possible to relate random-conical data sets to one another prior to reconstruction; and (c) the foundation is laid for a 3-D variance estimation which utilizes the oversampling of 3-D angular space by projections in the random-conical reconstruction scheme.

Algorithms↗

PubMed1992

Magnetic resonance neurography.

We have made cross-sectional image "neurograms" in which peripheral nerve has a greater signal intensity than that of other tissue. Neurographic images of the rabbit forelimb were obtained using a spin-echo magnetic resonance imaging (MRI) technique that combines fat suppression and diffusion weighting. After fat suppression the nerve shows up in relative isolation and is brighter than the surrounding tissue due to its longer T2 relaxation time of approximately 50 ms compared to approximately 27 ms for muscle. The addition of pulsed gradients for diffusion weighting of the MR signal further enhances the intensity of the nerve signal relative to that of surrounding muscle tissue. The greater diffusional anisotropy of nerve tissue (D parallel/D perpendicular = 3.1) compared to that of muscle (D parallel/D perpendicular = 1.9) allows further enhancement of the nerve by a subtraction of two diffusion-weighted images, one with the gradients oriented parallel and one with the gradients oriented perpendicular to the nerve orientation. We show that by manipulation of the MRI parameters, either echo time or pulsed gradient strength, the nerves can be made to show up as the most intense feature. This verifies the feasibility of generating three-dimensional "neurographic" images, analogous to angiograms, but which demonstrate the peripheral nerve tracts in apparent isolation.

Adipose Tissue↗

PubMedDate not supplied

Magnetization-prepared MR angiography with fat suppression and venous saturation.

Magnetization-prepared magnetic resonance (MR) angiography (MPMRA) is an inflow-based two-dimensional (2D) imaging sequence in which a preparation phase precedes rapid image acquisition. For maximal blood/tissue contrast, an inversion-recovery preparation nulls signal from static tissue. If needed, a second inversion suppresses signal from fat. Fully magnetized blood flows in after the inversion pulse(s), providing high signal intensity. The centric phase-encoding order, which ensures that the initial contrast is reflected in the image set, requires the use of a modified venous saturation technique. The sequence is described and its performance assessed with regard to (a) depiction of in-plane flow, (b) fat suppression, and (c) venous saturation. Phantom and volunteer studies showed good performance in all three areas. MPMRA images, acquired in just 2-4 seconds per image, had a blood/tissue contrast-to-noise ratio nearly twice that of standard 2D time-of-flight MR angiograms, acquired in 5-7 seconds. The technique is promising for restless patients and in anatomic areas plagued by motion degradation.

Adipose Tissue↗

PubMed1992

Uses and limitations of spoiled gradient-refocused imaging in the evaluation of suspected intracranial tumors.

This article describes the use of a radiofrequency-spoiled gradient-recalled (SPGR) imaging pulse sequence in the evaluation of intracranial masses. This pulse sequence provides excellent anatomic detail with T1-weighted image contrast. Rapid, sequential, single-slice (two-dimensional) images of the brain can be obtained in patients who are unable to hold still for long periods of time. In addition, volumetric (three-dimensional) image data sets can be obtained that provide extremely thin (1- to 2-mm) sections of high detail and good signal-to-noise ratio for selected critical structures within the brain. Finally, because SPGR is also utilized for time-of-flight angiography, parenchymal information can be obtained simultaneously with cerebral blood vessel definition. One potential pitfall that the magnetic resonance radiologist must be aware of is the fact that, in many patients, the degree of contrast enhancement is greatly diminished on postgadolinium SPGR images compared with conventional spin-echo T1-weighted images. Comparison with a set of standard spin-echo postcontrast images or, potentially, the use of higher doses of gadolinium may solve this problem. In spite of this limitation, the selective utilization of SPGR imaging can yield additional useful information for evaluation and preoperative planning in patients with intracranial masses.

Brain Neoplasms↗

PubMed1992

The recording and analysis of EMG and jaw tracking. I. The recording procedure.

A computer-based system is described which records and analyses electromyographic (EMG) signals and tracking data for mandibular movements during function. EMG signals were obtained bilaterally from six muscles and a Sirognathograph monitored the position of the lower incisor teeth in three dimensions. Directed jaw movement tasks by subjects were monitored. These data were continuously sampled at a rate of 1 KHz per channel over an operator controlled period. Analysis of the data was performed off-line following the experiment. Individual chewing cycles associated with masticatory function were identified and the timing and dimensions of each chewing envelope established. Corresponding EMG values were also determined and displays of EMG and jaw movement were plotted on a common time base. An average chewing cycle and correlated EMG activity could also be displayed on a high resolution monitor and plotted for a hard copy to enable a comparison to be made between subjects with and without jaw dysfunction.

Analysis of Variance↗

PubMed1992

Reconstruction of the electrocardiogram during heart surgery.

Electrocardiograms (ECG) recorded during arrhythmia surgery are used for identification of arrhythmias of different morphology. However, the interpretation of an intraoperative ECG is difficult because some leads cannot be recorded and the signals of the remaining leads often differ from those of a preoperative recording because of the sternotomy. Therefore, a method for reconstruction of a complete intraoperative ECG, which resembles a preoperatively recorded ECG, was studied in 24 patients undergoing heart surgery. The reconstruction method involves calculating coefficients for a transformation matrix, using a preoperative ECG recording and a first intraoperative ECG recording. Once this matrix has been established, further intraoperative recordings can be transformed into an ECG which strongly resembles a preoperative ECG. The correlation between reconstructed intraoperative leads and the corresponding preoperative leads was high in the leads Vx and Vy (median correlation coefficient 0.98 and 0.97) and slightly smaller in lead Vz (0.94). Further studies will prove if the method can be useful in arrhythmia surgery.

Aged↗

PubMed1992

Focal liver lesions: MR imaging with Mn-DPDP--initial clinical results in 40 patients.

Manganese (II) N,N'-dipyridoxylethylenediamine-N,N'-diacetate-5,5'-bis(phosphate) (DPDP) was evaluated as a contrast agent for magnetic resonance (MR) imaging (1.5 T) of focal liver lesions in 40 patients. Doses of 5 and 10 mumol/kg were administered intravenously. Mn-DPDP-enhanced T1-weighted images were compared quantitatively and subjectively with standard T1- and T2-weighted nonenhanced images. Use of Mn-DPDP resulted in a statistically significant increase in signal intensity of liver parenchyma in T1-weighted images at both doses. No enhancement was seen in metastases, cholangiocarcinomas, or lymphomas, while all hepatocellular carcinomas were enhanced. Enhancement was seen in focal nodular hyperplasia and in regenerative nodules. The lesion-to-liver contrast in Mn-DPDP-enhanced gradient-recalled-echo images was superior to that of all precontrast images (P less than .01). The number of nonenhancing malignant liver lesions detected in spin-echo (SE) images was increased (272 in T2-weighted SE images vs 390 in T1-weighted Mn-DPDP-enhanced SE images). Image interpretation (eg, visualization and demarcation of the lesions) was markedly better in Mn-DPDP-enhanced images than in all precontrast images (P less than .001).

Adult↗

PubMedDate not supplied

Dynamic analysis of drug action on in vitro reconstituted thyroid follicle by microinjection of tracer molecules and videomicroscopy.

Thyroid cells isolated from the gland by trypsinization are capable in culture of reconstituting histiotypic structures, the thyroid follicles. This morphological differentiation requires the presence of the main thyroid regulator; thyrotropin. We have analyzed some structural and functional aspects of in vitro reconstituted thyroid follicles (RTF) using microinjection of fluorescent probes and videomicroscopy. This experimental approach allowed to visualize biological processes and actions of drugs, signalling factors, etc. in living cells. We describe here some examples of what can be studied with this powerful still-undervalued method. Microinjection of a cell-impermeant fluorescent probe of either high or low molecular mass into the lumen of RTF allowed to check the tightness of this compartment and therefore to analyze the control of tight junctions assembly. A small cell-impermeant probe like Lucifer Yellow microinjected into a cell was used to demonstrate and then to study the regulation of cell to cell communication via gap junctions. The presence of calcium in the lumen of RTF was detected by microinjection of a properly designed probe: Calcium Green which becomes fluorescent in the presence of the ligand. The lumen to cell transport or endocytosis of thyroglobulin, the thyroid prohormone, which is stored into the lumen of the follicles, is currently studied by microinjection of TRITC-labeled thyroglobulin. Coupled to image processing and videorecorder systems, kinetic analysis and quantitative measurements can be performed.

Animals↗

PubMed1992

A novel principle for quantitation of fast intracellular calcium changes using Fura-2 and a modified image processing system--applications in studies of neutrophil motility and phagocytosis.

A new principle is described for imaging intracellular free calcium [Ca2+]i changes in single, living cells utilizing the fluorescent probe Fura-2. It is based upon video color mixing in real time and allows high-speed visualization, at maximum image resolution, of [Ca2+]i changes without digital image ratioing. The epifluorescence images produced by 340 and 380 nm excitations are stored in two memory buffers of a personal computer-based image processing system. Two video signals are generated independently from each buffer and connected to the red and green inputs of a video display. An image is this way created, in which [Ca2+]i shows up as a specific hue, whereas changes in dye concentration, light intensity, cell thickness show up as variations in brightness of the imaged cells. The method has advantages over conventional ratio imaging, notably simplicity and speed, since no calculations are made. Yet it can be combined with traditional digital image processing. The imaging technique allows monitoring of [Ca2+]i changes in rapidly moving cells, like neutrophils. It is demonstrated that during random locomotion on serum-coated glass surfaces, [Ca2+]i levels appeared to oscillate and that the frequency of the oscillations are related to locomotive activity. Furthermore, in Ca2+ free medium, the cells continue to move and phagocytose in the presence of Ca2+ ionophore (ionomycin) and 2 mM EGTA. In the presence of 1 mM extracellular Ca2+, ionomycin-treated cells were not able to move or phagocytose.

Calcium↗

PubMed1992

Poly(A) RNA codistribution with microfilaments: evaluation by in situ hybridization and quantitative digital imaging microscopy.

The distribution of poly(A) RNA has been visualized in single cells using high-resolution fluorescent in situ hybridization. Digital imaging microscopy was used to quantitate the signal in various cellular compartments. Most of the poly(A) signal remained associated with the cellular filament systems after solubilization of membranes with Triton, dissociation of ribosomes with puromycin, and digestion of non-poly(A) RNA with ribonuclease A and T1. The actin filaments were shown to be the predominant cellular structural elements associating with the poly(A) because low doses of cytochalasin released about two-thirds of the poly(A). An approach to assess the extent of colocalization of two images was devised using in situ hybridization to poly(A) in combination with probes for ribosomes, membranes, or F-actin. Digital imaging microscopy showed that most poly(A) spatially distributes most significantly with ribosomes, slightly less with F-actin, and least of all with membranes. The results suggest a mechanism for anchoring (and perhaps moving) much of the cellular mRNA utilizing the interaction between actin filaments and poly(A).

Actin Cytoskeleton↗

PubMed1992

[Clinical potentialities and limitations of two- and three-dimensional time-of-flight MR angiography in the diagnosis of carotid stenosis].

For diagnosing a carotid artery stenosis, 26 patients underwent MR angiography. A spin echo (SE) sequence with presaturation on which the flowing blood appears with low signal intensity was applied, as well as a two- and three-dimensional gradient echo (GRE) sequence with flow compensation, showing blood flow with high signal intensity. Subsequently, projection angiograms were made from the MR images with a maximum intensity projection algorithm. Degree, length and localisation of a carotid artery stenosis were reviewed. To find out the clinical usability, the results of the MR angiograms were compared subsequently with the findings of intravenous digital subtraction angiography (i.v. DSA). In comparison with DSA a correlation in the degree of stenosis was noticed in 42 out of 48 SE-, in 39 out of 52 2-D GRE and in 41 out of 48 3-D GRE sequence angiograms. The length of 34 moderate and severe stenoses, demonstrated by DSA, was overestimated 5 times with the SE-sequence, 9 times with the 3-D and 23 times with the 2-D GRE sequence. MR angiography with a 3-D GRE sequence is suitable for screening for carotid artery stenosis. In cases of severe stenosis an SE sequence should be performed for more precise delineation of the stenotic lesion.

Aged↗

PubMed1992

An NMR phased array for human cardiac 31P spectroscopy.

A four-coil phased-array 31P NMR receiver was designed and tested for human cardiac applications, to determine whether the combination of relatively high signal-to-noise ratio (SNR) and large field of view produced in 1H imaging is also realized for in vivo 31P spectroscopy. Spectra were acquired in parallel from an array of four overlapping 6.5-cm surface coils using one- and two-dimensional phase-encoding pulse sequences and were optimally combined to yield composite spectroscopic images. The phased array was found to generate useful 31P spectra from a 2.5-fold wider lateral region around the anterior myocardium than a single receiver of the same size as the array elements, with no increase in imaging time. In addition, the sensitive depth was increased by up to 2 cm over that of a single coil. Spectra could be acquired in roughly 15 min from a region extending to the middle of the heart, with voxel sizes of 2 x 2 x 4 cm3. For the average heart voxel, the SNR of the combined spectrum was higher than that of the best spectrum from any one coil in the array by 30%, with some voxels showing an increase as high as 60%.

Heart↗

PubMed1992

Magnetic resonance angiography techniques.

After a radio frequency pulse, the decay of the magnetic resonance (MR) signal is described by two relaxation processes, T1 and T2. T1 describes the rate at which the magnetization realigns itself along the external magnetic field direction (ML), and T2 describes the rate of decay of the magnetization component along the transverse axis (MT). Magnetic resonance angiography (MRA) sequences have been developed that encode flow as changes in the apparent T1 or T2 of the moving blood relative to stationary tissues. MRA sequences typically use either time-of-flight (TOF) techniques to encode T1 or phase-contrast techniques to encode T2. TOF techniques encode flow as an apparent T1 shortening through the wash-in of fully relaxed blood from outside the image volume. The shorter T1 produces an enhancement of vascular structures relative to stationary tissues. TOF methods may use either sequential two-dimensional, three-dimensional, or multi-slab three-dimensional imaging sequences to produce a three-dimensional MRA data set. Phase-contrast methods use additional magnetic field gradients to encode flow as shifts in the phase of MT. Both TOF and phase-contrast methods use maximum intensity projection (MIP) images displayed in a cine format to aid in the visualization of three-dimensional vascular structures.

Animals↗

PubMed1992

Quantification of inter- and intra-nuclear variation of fluorescence in situ hybridization signals.

This study aims at the quantification of specific DNA sequences by using fluorescence in situ hybridization (ISH) and digital imaging microscopy. The cytochemical and cytometric aspects of a quantitative ISH procedure were investigated, using human peripheral blood lymphocyte interphase nuclei and probes detecting high copy number target sequences as a model system. These chromosome-specific probes were labeled with biotin, digoxigenin, or fluorescein. Quantification of the fluorescence ISH signals was performed using an epifluorescence microscope equipped with a multi-wavelength illuminator, and a cooled charge coupled device (CCD) camera. Specific image analysis programs were developed for the segmentation and analysis of the images provided by ISH. The fluorescence intensity distributions of the ISH spots showed large internuclear variation (CVs up to 65%) for the probes used. The variation in intensity was found to be independent of the probe, the type of labeling, and the type of immunocytochemical detection used. Variation in intensity was not caused primarily by the immunocytochemical detection method, since directly fluorescein-labeled probes showed similar internuclear variation. Furthermore, it was found that different white blood cell types, which harbor different degrees of compactness of the nuclear chromatin, showed the same variation. The intra-nuclear variation in intensity of the ISH spots on the two chromosome homologs within one nucleus was significantly smaller (approximately 20%) than the inter-nuclear variation, probably due to more constant local hybridization conditions. Due to the relatively small intranuclear variation, copy number polymorphisms of the satellite DNA sequence on chromosome 1 could readily be quantified.(ABSTRACT TRUNCATED AT 250 WORDS)

Analog-Digital Conversion↗

PubMed1992

Release of intracellular calcium and modulation of membrane currents by caffeine in bull-frog sympathetic neurones.

1. Calcium release and sequestration were studied in whole-cell voltage-clamped bull-frog sympathetic neurones by image analysis of Fura-2 signals. 2. Application of caffeine (10 mM) to cells voltage clamped at -38 mV caused a rapid increase in intracellular calcium concentration ([Ca2+]i) to a mean value of 352 +/- 33 nM, which activated an outward current. In the continued presence of caffeine the rise in [Ca2+]i slowly declined to a sustained plateau of 196 +/- 20 nM (112 nM above control levels), while the outward current rapidly decayed. Peak calcium release was highest at the edge of the cell. 3. The caffeine-evoked intracellular calcium increase was reduced by two inhibitors of calcium-induced calcium release, ryanodine and procaine. The residual non-suppressible increase in [Ca2+]i may indicate that caffeine can release calcium from two pharmacologically distinct intracellular stores. 4. Inhibition of the caffeine-evoked release of calcium by ryanodine was both concentration and 'use dependent' so that the full inhibitory effect was only observed when caffeine was applied for the second time in the presence of ryanodine. In contrast, the action of procaine did not show any 'use dependence' and unlike ryanodine was fully reversible. 5. The outward current was sensitive to blockers of the large conductance calcium-activated potassium current, Ic. Analysis of variance from this current indicated that it arose at least partly from summation of spontaneous miniature outward currents. 6. The magnitude and duration of calcium release by caffeine was dependent on the resting level of intracellular calcium and the caffeine exposure time. This, together with the pharmacology of the release, suggests that caffeine increases intracellular calcium by sensitizing calcium-induced calcium release. 7. The evoked [Ca2+]i increase was enhanced in amplitude by intracellular application of Ruthenium Red. This effect was mimicked by extracellular application of the mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP) but not by internal application of FCCP or other inhibitors of mitochondrial Ca2+ uptake. This suggests that the evoked increase in [Ca2+]i is predominantly buffered by a Ruthenium Red-sensitive sequestration process which is not mitochondrial.

Animals↗

PubMedDate not supplied

MR imaging with spatially variable resolution.

In some situations it may be advantageous to produce "locally focused" magnetic resonance images that have nonuniform spatial resolution matching the expected local rate of spatial variation in the object. Because such an image has fewer pixels than a conventional image with uniformly high resolution, it can be reconstructed from fewer signals, acquired in less time. This can be done by using a highly convergent representation of the image as a sum of orthonormal functions with slow (fast) spatial variation in relatively homogeneous (heterogeneous) parts of the object. Since this series is shorter than a conventional truncated Fourier series, its terms can be calculated from a subset of the usual array of phase-encoded signals. The optimal choice of these phase encodings, which are usually scattered nonuniformly in k space, results in minimization of noise in the reconstructed image. The technique is illustrated by applying it to simulated data and to data from images of phantoms.

Computer Simulation↗

PubMed1977

The role of the digital computer in pediatric cardiology.

A digital computer system is described which allows the real-time processing of all physiological signals obtained during a heart catheterization procedure and which makes all relevant results and informations available immediately during the investigation. In addition, special electronic units and programs have been developed in our institution for the automated extraction of morphological criteria from biplane angiocardiograms. Thereby right and left ventricular volume, shape and contraction pattern can be quantitated and used to characterize the performance of the heart as muscle and pump in physical terms. Recently, complete digital processing of videoangiocardiograms has been achieved in a stroboscopic mode, each videofield in real time. Application of image enhancement, subtraction, integration and restoration techniques leads to a fundamentally improved angiocardiographic image quality for a given amount of injected contrast material. Based on eight years of experience with digital computer application in pediatric cardiology, computer technologies are considered likely to become the method of choice in the future.

Angiocardiography↗

PubMed1975

Quantitative colour-ultrasonography by means of a computer aided simultaneous tomogram.

In this newly developed method, the ultrasonic diagnosis has become quantitative. The data will remain quantitative so far as an image storage tube is used. With the conventional storage type CRT, it is impossible to memorize the reflected echo signals of different strength levels. But with our method, it becomes possible to memorize the images according to individual levels. By the direct digitalization of the ultrasonic echo images, more information is obtained from the reflected echo signals. By processing the original information with a computer, more accurate diagnosis is possible. It is hoped and believed, that this computer aided ultrasonic examination, with single scan display in colour or black and white, will further promote ultrasonic diagnosis in future.

Color↗