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Biomedical subjects

J M Thijssen

Publications and source records attributed to J M Thijssen.

At least 19 recordsLinked to original sources

Effects of tissue processing techniques in acoustical (1.2 GHz) and light microscopy.

In this study the influence of various tissue processing and staining techniques on the acoustical properties of liver tissue was investigated. A qualitative study was performed using ultrasound attenuation as the imaged parameter of a combined optical/acoustical microscope with a 1.2 GHz transducer. Images were made of three sets of adjacent liver sections (6 microns in thickness) which were prepared in ten different ways: fixed by alcohol or formalin; stained by hematoxylin-eosin (HE), toluidine blue (TB) or non-stained; sectioned by a cryostat or by a paraffin microtome. It was concluded that the images obtained from cryostat sections were of much higher quality than those from paraffin sections. Images obtained from sections that were sectioned while embedded in paraffin displayed no detail at all. No consistent effect was noticed with respect to staining by HE or TB. Alcohol fixed sections gave more detailed images than formalin fixed sections. Formalin fixation in combination with cryostat sectioning yielded many cytoplasmic vacuoles.

Alcohols

In vitro classification of gallstones by quantitative echography.

Gallstones (n = 20) were classified by dual energy Computer Tomography (CT) into three main classes: pure cholesterol stones (I), combination stones (II) and calcium stones (III). Further subclassification was possible by using morphological criteria. The acoustic measurements that were performed were measurements of the velocity of sound, the attenuation coefficient slope with frequency and intercept at 4.5 MHz, the attenuation coefficient slope per unit of time, the backscattering characteristics, and the appearance of B-mode echograms. The velocity of sound in calcium stones (c = 1695 +/- 107 m/s) was distinctly lower than in those containing some cholesterol (c > 2000 m/s). The attenuation coefficient slope ranged from 4.3 to 16.2 dB/cm MHz, the 4.5 MHz intercept from 21 to 66.2 dB/cm. The lowest values were found for the pure cholesterol stones (class IA), the highest values for subclass IIB (combination stones with shell). The attenuation coefficient slope per unit of time was distinctly lower (< 0.50 dB/microseconds MHz) for the cholesterol stones than for the combination and calcium stones (>0.64 dB/microseconds MHz). The backscattering spectrum was approximated by a straight line fit, and the slope for the cholesterol stones was lower than for the combination and calcium stones (<0.75 dB/MHz vs. >1.0 dB/MHz, respectively). The latter two parameters were assessed by in vivo applicable methods. The front echo level was found to be more than 5 dB higher for class IIB as compared to the other classes, while the spectral backscatter level at 4.5 MHz was considerably higher for both classes IIB and III. The B-mode echograms showed that a strong front echo in combination with a strong attenuation of the remaining echo signals was mainly found for stones of classes IIB and III. A significant difference between the group of stones that are suitable for lithotripsy and dissolution treatment (classes IA, IB and IIA) and the nontreatable calcium-containing stones (classes IIB and III) was found for the velocity of sound (p <0.01), the attenuation coefficient slope per time unit (p <0.10), the slope of the backscattering spectrum (p <0.05) and the 4.5 MHz intercept (p <0.01). B-mode classification yielded no complete distinction of these two groups of stones. It can be concluded that in vivo assessment of quantitative characteristics (front reflection, backscattering characteristics, attenuation coefficient slope per unit of time) in combination with the B-mode characteristics might be useful for in vivo gallstone classification.

Cholelithiasis

Echographic differentiation of intraocular melanomas by computer analysis.

The aim of this study was to assess the differentiation of histologically different types of choroidal melanomas by means of clinical and quantitative acoustic/texture parameters of echograms. Clinical parameters were graded by morphological, kinetic and quantitative statements about the A- and B-mode echograms by a skilled diagnostician. Acoustic/texture parameters were obtained by processing and analysing radio frequency, AM-demodulated and FM-demodulated echograms. The latter data were preprocessed to remove influence induced by beam diffraction and focusing. The best set of four clinical echographic parameters enabled a retrospective classification of spindle type melanomas vs. mixed+epithelioid type melanomas with an accuracy of 77% (area under the ROC-curve of 86%). The discriminant analysis performed with the best four acoustic/texture parameters (n = 30) yielded a sensitivity of 89%, a specificity of 92%, and an area under the ROC-curve which corresponds to a probability of correct classification of 96.6%. When using these data prospectively to classify tumours of unknown cell type (n = 21), this classification could be performed in 86% of cases.

Algorithms

B-mode echography of choroidal melanoma; echographic and histological aspects of choroidal excavation.

The B-mode images of 32 patients affected with a malignant melanoma of the choroid were studied, in particular with respect to choroidal excavation. The degree of visibility of choroidal excavation at the B-scan was compared with the thickness of the choroid, scleral invasion, vascularisation, retinal detachment and the reflectivity of the tumour. None of these histological and echographical features appeared to be significantly related to the degree of the visibility of choroidal excavation. Also the condition of the choroid was studied. At examination of all the histological sections no choroid was found between tumour and sclera. This implies that choroidal excavation should be visible at echograms in every case instead of only in 75% of the cases as was found for our series.

Choroid

Variability of quantitative echographic parameters of the liver: intra- and interindividual spread, temporal- and age-related effects.

The values of acoustic and image texture parameters were estimated from liver scans of healthy subjects. The values were obtained after appropriate preprocessing of the radio frequency echograms by an on-line computerized system. The preprocessing comprised a correction for the Time Gain Compensation (TGC), the beam diffraction and the frequency dependent attenuation in the Region of Interest (ROI). The intra- and interindividual variability of the parameter values appeared to be of the same order of magnitude, but significantly larger than the variability assessed by measurements of a homogeneously scattering tissue mimicking phantom. Significant temporal effects were found for all the parameters, which consistently occurred during the morning. These results are discussed in relation to the circadian rhythm of the glycogen content and of the hepatic circulation. All the parameters appeared to be significantly correlated to age. The slope of the regression ranged from 3.6% per decade (attenuation coefficient) to 7.6% per decade (mean echo-level). A tentative explanation to these results is presented: the increased stiffness of hepatic vasculature with age.

Adult

Influence of histochemical preparation on acoustic parameters of liver tissue: a 5-MHz study.

In this study the influence of various histological techniques on the acoustic parameters of liver tissue was investigated. Radiofrequency (RF) echographic data were obtained in vitro from 21 liver samples taken from 8 white New Zealander rabbits. The samples were measured in four different subsequent histological tissue processing conditions (freshly excised, 4% buffered formalin fixed, after it went through a paraffin cycle and after staining with hematoxylin and eosin). The acoustic parameters that were obtained from the rf data were velocity of sound, slope of the attenuation coefficient versus frequency between 1.9 and 6.9 MHz, attenuation coefficient at 4.4 MHz, slope of the backscattering spectrum between 1.9 and 6.9 MHz, and intercept of the backscattering spectrum. It was found that fixation by formalin preserves the acoustic properties of the tissue to a reasonable extent. Embedding in paraffin and deparaffinizing induces large changes in the acoustic properties of the tissue. As an alternative, freezing prior to cutting, rather than the paraffin cycle, was investigated also in 10 liver samples obtained from 4 New Zealander rabbits. This method produced no significant changes of the acoustic parameters and should therefore be preferred in acoustic microscopy.

Acoustics

Echographic differentiation of histological types of intraocular melanoma.

The aim of this study was to assess the differentiation of histologically different types of choroidal melanomas by means of clinical and quantitative acoustic/texture parameters of echograms. Clinical parameters were graded by morphological, kinetic and quantitative statements about the A- and B-mode echograms by a skilled diagnostician. Acoustic/texture parameters were obtained by processing and analysing radio frequency, AM-demodulated and FM-demodulated echograms. The data were preprocessed to remove influences induced by beam diffraction and focusing. The correlations between the clinical parameters were lower than 0.5 but significant in a few cases. The best set of four clinical echographic parameters enabled a retrospective classification of spindle type melanomas vs. mixed + epithelioid type melanomas with an accuracy of 77% (area under the ROC-curve of 86%). These figures do not enable, however, a prospective diagnosis. The correlations of the clinical parameters with the acoustic/texture parameters were investigated. The mutual correlations between the latter parameters were also assessed, the most significant (0.70) being between the speckle size and the slope of the linearized backscattering spectrum. The discriminant analysis performed with the best four acoustic/texture parameters (n = 30) yielded a sensitivity of 89%, a specificity of 92% and an area under the ROC-curve corresponding to a probability of correct classification of 96.6%. When using these data prospectively to classify tumours of unknown cell type (n = 21), this classification could be performed in 86% of cases.

Algorithms

Ultrasound attenuation and texture analysis of diffuse liver disease: methods and preliminary results.

A study was performed to find and test quantitative methods of analysing echographic signals for the differentiation of diffuse liver diseases. An on-line data acquisition system was used to acquire radiofrequency (RF) echo signals from volunteers and patients. Several methods to estimate the frequency-dependent attenuation coefficient were evaluated, in which a correction for the frequency and depth-dependent diffraction and focusing effects caused by the sound beam was applied. Using the estimated value of the attenuation coefficient the RF signals themselves were corrected to remove the depth dependencies caused by the sound beam and by the frequency-dependent attenuation. After this preprocessing the envelope of the corrected RF signals was calculated and B-mode images were reconstructed. The texture was analysed in the axial direction by first- and second-order statistical methods. The accuracy and precision of the attenuation methods were assessed by using computer simulated RF signals and RF data obtained from a tissue-mimicking phantom. The phantom measurements were also used to test the performance of the methods to correct for the depth dependencies. The echograms of 163 persons, both volunteers and patients suffering from a diffuse liver disease (cirrhosis, hepatitis, haemochromatosis), were recorded. The mutual correlations between the estimated parameters were used to preselect parameters contributing independent information, and which can subsequently be used in a discriminant analysis to differentiate between the various diseased conditions.

Adult

Ultrasonic differentiation of intraocular melanomas: parameters and estimation methods.

In this study, the estimation of ultrasound parameters is evaluated for in vivo differentiation of intraocular melanomas. For this purpose, both tissue and image parameters of the ultrasound signal are considered. These parameters comprised, respectively, the frequency dependent attenuation and backscattering coefficient of the melanoma tissue, and the first and second-order statistics of the amplitude-modulated and phase-derivative images of the melanomas. A diffraction correction procedure has been applied prior to the estimation of the parameters to correct the ultrasound signals for the echographic equipment used and for the various distances of the region-of-interest to the transducer. In addition, a pre-processing to select a homogeneous region from the tumours was implemented to obtain consistent estimates of the ultrasound parameters, because the accuracy and the precision of the parameters would be greatly reduced by the inhomogeneity of the melanoma tissue. The estimation methods are evaluated by means of the accuracy and precision of the parameters estimated from simulated ultrasound data and data obtained from a tissue-mimicking phantom. The mutual correlations of the parameters are discussed for the ultrasound data obtained from the melanomas. This study enabled a preselection of the independent ultrasound parameters that could be used in a discriminant analysis to perform a differentiation of intraocular melanomas. The sensitivity and specificity of differentiating spindle cell type from mixed-epitheloid meleanomas were 92 and 89 percent, respectively.

Choroid Neoplasms

Improvement of lesion detection by echographic image processing: signal-to-noise-ratio imaging.

Signal-to-noise-ratio (SNR) images were constructed by a sliding window technique. Theoretical derivations of the first order statistical properties of SNR images are given for the case of intensity coding of the original images for low number densities and for the limit case of fully developed speckle. The original images were obtained by realistic simulations of backscattering by homogeneous media, while using a range of number densities of the scatterers (isotropic scattering). The SNR imaging method is illustrated by results obtained from simulated scattering media containing a lesion that is contrasting in reflectivity level or in number density of the scatterers. A considerable improvement in lesion detection is obtained if the primary difference from the background speckle is due to a different number density of the scatterers within the lesion.

Image Processing, Computer-Assisted

A simulation study of echographic imaging of diffuse and structurally scattering media.

Realistic simulations of echographic image formation were performed. The simulations were based on a focussed single element transducer of 3.5 MHz and linear scanning. The tissue model scanned was composed of a homogeneous, nonattenuating, medium containing point-like scatterers. The scatterers were distributed in space in two different arrangements: randomly with a fixed number density of 7500 per cubic centimeter and regularly in a cubic matrix with a characteristic distance of 1 mm. These two populations of scatterers yielded the diffuse and the structural scattering component, respectively. The B-mode texture was assessed by first and second order statistical parameters. The effects of various modifications of the matrix, i.e., the relative scattering strength, the orientation and the position uncertainty of the scatterers, on the statistical parameters were systematically investigated. Increasing the relative scattering strength yields a monotonic increase of the mean grey level, the ratio of structural over diffuse scattering intensity and a decrease of the signal-to-noise ratio and the autocorrelation lengths. Increasing the position uncertainty to 20 percent reduces these effects practically to zero, except for the lateral ACF's. Rotation of the matrix with respect to the beam axis induces a periodicity in all statistical parameters, which is symmetric around 45 degrees. Again, the effects become statistically insignificant at a position uncertainty of 20 percent, with the exception of length of the lateral ACF. While assuming that in a single clinical B-mode echogram of the liver many orientations of the matrix will be simultaneously present, the B-mode lines of images obtained for a range of orientations were taken together and the overall statistical parameters calculated. The mean, the signal-to-noise ratio of the echo amplitude, SNRA, and the axial and lateral lengths of the ACF for the multirotation condition are significantly different from both the diffuse and the zero degree orientation. The structural backscattering intensity cannot be estimated any more with a reasonable precision. The structural separation distance, however, can still be assessed at a 10 percent level of the position uncertainty, even in the multirotation case and is therefore a robust clinical parameter.

Computer Simulation

Phase-derivative imaging. III: Theoretical derivation of first and second order statistics.

The echo signal obtained from a homogenous and isotropically scattering medium can be described as a Poisson time series which is convolved with the transmission pulse of the transducer. The probability density function (pdf) of this signal approximates to a Gaussian pdf for narrowband pulse waveform. Methods to derive the phase-derivative (PD) signal from the complex envelope and the preenvelope of the echo signal are described. The first order pdf of the PD asymptotically becomes a Gaussian pdf by smoothing. Since the rectified PD is employed to obtain 2-dimensional grey scale images, the first order pdf as well as the signal-to-noise ratio (SNR) of this signal are also derived. The rectified PD is further smoothed by a cosine time window prior to the imaging. The SNR and the autocorrelation function (in the axial direction) of this latter signal can be derived under the assumption of a Gaussian spectrum of the transmission pulse. These first and second order characteristics of the PD images are calculated for the conditions employed in simulations and experiments reported previously and are quantitatively compared to the values obtained from these.

Biophysical Phenomena

Texture in tissue echograms. Speckle or information?

Models of biological tissues are described in terms of acoustic parameters and of structure. Beam formation is discussed for continuous wave and pulsed modes of transducer operation and the concept of the point spread function (PSF) is introduced. The PSF is equivalent to the resolution cell, or the sampling volume, of echographic equipment. The generation of echograms from parenchymal tissues is described in terms of speckle formation due to interference at reception on the transducer. The speckle dimensions are quantitatively compared to the sampling volume of the employed transducer. It is shown that for fully developed speckle the tissue characteristics are exclusively reflected in the mean echolevel and not in the speckle size. The speckle size is, however, greatly dependent on the bandwidth, the frequency, and the geometry of the employed transducer. The attenuation by the insonated tissue yields a depth-dependent increase of mainly the lateral speckle size, in addition to the depth dependence caused by the beam formation. If the number density of scattering sites within the tissue is relatively low, the speckle characteristics are dependent on this density and, hence, tissue characterization is feasible if these characteristics are analyzed by statistical methods. These methods are gray level histogram analysis and the estimation of the autocorrelation function, ie, first and second order statistics, respectively. Structural order in tissues can be quantified by autocorrelation analysis and clinical studies on diffuse liver diseases support this conclusion. The effects of pre- and postprocessing on the detectability of focal lesions are outlined. The impact of multifocus systems and of the acquisition of radio frequency echograms on further developments of clinical echography is discussed.

Acoustics

In vivo ultrasound backscattering estimation for tumour diagnosis: an animal study.

In this study the efficacy of a quantitative analysis of backscattered ultrasound for the differential diagnosis of intraocular tumours is tested. The data acquisition was performed with an on-line acquisition and processing system for in vivo work. The backscattering spectra were corrected for the beam effects (diffraction and focussing). The measurements were taken from an animal model (rabbit) in which a Greene's amelanotic melanoma was implanted in the anterior chamber of the eye. The special histologic arrangement of this tumour makes it optimally suited to an assessment of the correlation between histology and backscattering cross section. Various backscattering models were considered theoretically and the choice for a practical testing was motivated on the properties of the observed backscattering spectra. We conclude that the backscattering model based on an "inhomogeneous continuum" with a cylindrical Gaussian autocorrelation function fits the data optimally. The relatively low correlation of the backscattering cross-sections to histology cannot yet be fully explained. The range of acoustic scatterer sizes, however, corresponds quite well to the dimension of observed and quantified histologic structures.

Animals

Ultrasonic tissue characterisation and echographic imaging.

Acoustic models of soft biological tissues are discussed. The methods to derive acoustic bulk parameters, i.e. attenuation coefficient and backscattering coefficient are outlined. When applying a specific backscattering model the effective dimension of the scattering sites within a tissue can be estimated. The texture of echographic images is defined in terms of speckle and it is discussed in which way the texture is influenced by the beam formation and the volume density of the scatterers. Structural characteristics of tissues are discussed and the method to estimate a structural dimension is indicated. Recent developments are parametric imaging and image processing.

Acoustics

Phase-derivative imaging. II: Effects of beam diffraction and scatterer density.

The potential of using the phase derivative (PD) of the radio-frequency echograms for producing 2-dimensional grey scale images was further investigated. The PD images were produced by five different algorithms, which according to the results described in the companion paper yield PD images dominated by the amplitude (envelope, ENV), mixed PD-AM images and pure PD images. These images are termed according to their algorithms: ZCS, zero crossing counter with squelch; ASS, analytic signal with squelch; ASW, analytic signal with Wiener kernel; UNP, unwrapped phase; and SAS, smoothed analytic signal. The rf data were obtained from simulations and from experiments with a tissue mimicking phantom. PD images were analysed by calculation of the first and second order grey level statistics: mean level, signal-to-noise ratio (SNR) and the full width at half maximum (FWHM) of the autocovariance functions (ACVF). These parameters were systematically investigated for a range of depths with respect to the transducer and a range of scatterer densities of the insonated medium. The UNP and SAS images do not suffer much from the diffraction effect but do not display much information about the scatterer density either. The ASW and ASS images qualitatively display beam diffraction effects similar to those of the AM images, with the exception of the mean value which is at the minimum in the focus, where the AM yields a sharp maximum at that depth. The mean and the SNR of the ASW and ASS images increase with increasing scatterer density and saturate at a density of 5000 cm-3. The mean value of the envelope, however, displays a square root dependency over the whole range. The axial and lateral FWHM of the ACVF of the UNP and SAS methods are not significantly dependent on the scatterer density and decrease with increasing density for the ASW and ASS images, as was observed in the envelope images. It may be concluded that ASW and ASS methods produce PD grey scale images which are equally well suited for the diagnosis of diffuse diseases of parenchymal tissues as conventional AM images. The smaller "speckle" size of the ASW images might be advantageous, for the detection of focal lesions, but the lesion contrast is found to be much lower than for the ENV.

Computer Simulation

Diagnostic ultrasound equipment. Safety and dosimetry.

A survey of present opinions on the safety of diagnostic ultrasound applications is presented together with some data on output levels. Physical mechanisms involved in potential adverse effects are described. Labelling requirements of the equipment are defined and specified. These requirements are contained in a recent proposal of the Netherlands' Committee on the Safety of Ultrasound.

Acoustics

Gray level transforms and lesion detectability in echographic images.

In search of the optimal display of echographic information for the detection of focal lesions, a systematic study was performed considering a wide range of gray level transforms (i.e., lookup tables). This range comprised power functions of the echo envelope signal (1/8 less than or equal to n less than or equal to 8), power functions of the logarithmic transform and a sigmoid function. The implications of the transforms on the first order statistics (histogram, "point signal-to-noise ratio" SNRp) and on the second order statistics (autocorrelation function) could be derived both analytically, and from the analysis of simulated and experimentally obtained echograms of homogeneously scattering tissue models. These results were employed to estimate the lesion signal-to-noise ratio SNRl, which specifies the detectability of a lesion by an ideal observer. It was found, both theoretically and practically, that the intensity display corresponds to the optimal transform (i.e., n = 2) for a low contrast lesion. When the data were first logarithmically compressed, the lesion SNR appeared to increase with increasing power (1/8 less than or equal to n less than or equal to 8). A logarithmic transform followed by a sigmoid compression did not produce much improvement. These effects of gray level transforms on the SNRl were shown to be relatively small, with the exception of powers n greater than 2 when applied to linear (i.e. amplitude) data. In the case of high lesion contrast, the sequence of log compression, followed by a square law produced the optimum SNRl. This sequence is equivalent to the processing within echographic equipment, where the TV monitor has a gamma of the order of 2.

Computer Simulation