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At least 91 records · Page 5Linked to original sources

Scanning acoustic microscopy visualizes cytomechanical responses to cytochalasin D.

The reactions of XTH-2 cells (line derived from Xenopus laevis tadpole hearts) to cytochalasin D (CD) were followed using scanning acoustic microscopy (SAM) at 0.9 GHz, fluorescence and electron microscopy. The first reaction to CD which can be detected by SAM is a loss of image contrast, indicating a decrease in acoustic impedance of about 30%. Based on structural changes revealed by staining of actin with TRITC-phalloidin, and taking theoretical considerations into account, a relationship between impedance decrease and tension in the actin fibrillar system is deduced.

Actins↗

Propagation speed of sound assessment in the layers of the guinea-pig esophagus in vitro by means of acoustic microscopy.

The study's purpose was to evaluate the propagation speed of sound in the tissue layers of the esophagus at various mechanical loadings. Scanning laser acoustics microscopy was applied for the estimation of the propagation speed in the mucosa-submucosa and muscle layers of guinea-pig esophagus in vitro (n = 26). The propagation speed in the esophagus was determined in the no-load state with all external forces removed, and in the distended and zero-stress states. The zero-stress state was obtained by cutting the esophageal rings radially. The propagation speed in the no-load state differed significantly (p < 0.001) between the muscle layer (median 1740, quartiles 1735-1746 m/s) and the mucosa (1607, 1605-1609 m/s). In the distended state the propagation speed in the muscle layer decreased significantly (p < 0.001) to 1673 (1666-1681) m/s while it did not change significantly in mucosa (1602, 1600-1607 m/s). When compared to the no-load state, the propagation speed in the zero-stress state in the muscle layers decreased to 1624 (1615-1636) m/s (p < 0.001) and in mucosa to 1584 (1566-1603) m/s (p < 0.001). In conclusion, the esophagus is a composite structure with heterogeneous propagation speed characteristics. Furthermore, the mechanical loading state must be considered in esophageal ultrasound studies.

Acoustics↗

Testing of metal-ceramic joint using scanning acoustic microscopy.

The objective of the investigation was to compare the results obtained from examination of titanium-porcelain joints by means of both scanning acoustic microscopy (C-SAM) and scanning electron microscopy (SEM). A dental porcelain (Ducera, Dental GmbH) mechanically compatible with titanium was fired to sand-blasted or electrolytically polished commercially pure titanium (grade 1) specimens. The firing was carried out in an ordinary dental furnace according to manufacturer's instructions. There was a good correlation between the C-SAM and SEM methods regarding the ability to detect air-filled defects in the porcelain/titanium interface. The results show that the C-SAM method, being non-destructive as well as time-and-money-saving, can be useful in the testing of metal-ceramic joints.

Dental Porcelain↗

Small intestine wall distribution of elastic stiffness measured with 500 MHz scanning acoustic microscopy.

Obtaining data relating intestinal mechanical properties and histology is a step towards the next level in the hierarchy of structure of living tissue, and may provide new insight into the mechanisms of intestinal function and disease such as obstruction. Due to lack of methodology, however, such data are currently sparse. Scanning acoustic microscopy (SAM) can measure the propagation speed of sound (C) and the acoustic impedance (Z) with micrometer resolution in tissue. By use of elementary theory of elasticity, the elastic stiffness (c11) can be computed from C and Z. We used 5-microm-thick transverse sections of ethanol treated guinea pig small intestine as the experimental model and measured the distribution of C and Z across the intestinal wall using SAM at 500 MHz. The individual layers mucosa, submucosa, and circular and longitudinal muscle were discerned with ease in the images and varied significantly with respect to both C and Z in most cases. The measured values (median values) of C ranged from 1550 to 1669 m s(-1), and Z ranged from 2.10 to 2.60 MPa s m(-1). c11 differed between all layers ranging from 3.25 to 4.27 GPa with the following sequence of magnitude: circular muscle >submucosa>mucosa>longitudinal muscle (p<0.001). In conclusion, we provided the first microscale mechanical data relating to the histological layers of the small intestine.

Acoustics↗

Evaluation of acetic acid-induced gastric ulcers in rats by scanning acoustic microscopy.

To clarify the mechanism of gastric ulcer recurrence, we studied acetic acid-induced gastric ulcers. The healing of these gastric ulcers was then studied for 4-21 weeks after the injection. Forty-eight ulcers were examined by macroscopic observation and scanning acoustic microscopy (SAM) and by histologic study of the ulcers. SAM has a higher radio frequency and provides more detailed information about the gastric wall than endoscopic ultrasonography (EUS). Thirteen healed and 35 non-healed gastric ulcers were observed by macroscopic observation over 4-21 weeks. SAM enabled us to observe the normal gastric wall of the rat in five different layers. The SAM echo patterns of the healed ulcers were all one pattern. On the other hand, we were able to classify the echo patterns of the non-healed ulcers into two different patterns; one pattern consisted of a high-echogenic region in the first and second layers with an underlying relatively high echogenic region in a horizontal pattern. The other pattern showed a poorly defined and spotted low-echogenic region in the first and second layers which fanned out toward the fifth layer. However, these two different ulcers could not be distinguished macroscopically. Histologic findings supported the results of the SAM study which showed one pattern in healed ulcers and two patterns in non-healed ulcers. In the non-healed ulcers the two different patterns may be ascribed to the proliferation of collagen fibers. SAM makes it possible to image the proliferation of collagen fibers and identify the two different types with an accuracy of 97%.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗

Quantification of atherosclerotic plaque composition in cholesterol-fed rabbits with 50-MHz acoustic microscopy.

To determine whether high-frequency ultrasound could distinguish normal from pathological vascular structure and to elucidate the determinants of ultrasonic backscatter in different layers of normal and atherosclerotic arteries, high-resolution acoustic microscopy at 50 MHz was used to characterize aortic plaque in six New Zealand White rabbits fed a 2% cholesterol diet for 3.5 months. Four rabbits were fed a standard diet for 3.5 months to provide normal control data. Segments of aortas were excised, fixed in formalin, opened longitudinally, and mounted flat for insonification. For each specimen, backscattered radio frequency (rf) data were acquired from 30 to 100 independent sites separated by 500 microns. Portions of rf data were gated from discrete layers of the vessel wall for computation of integrated backscatter. Results of histological and immunocytochemical analyses of vessel wall thickness and composition were compared with those of ultrasonic analysis. Normal aortas manifested prominent but homogeneous backscatter (average integrated backscatter, -28.5 +/- 2.9 dB) throughout the vessel wall, with no clear distinction between intimal and medial layers. The atherosclerotic aortas manifested substantially reduced integrated backscatter from the thickened intima (-47.5 +/- 3.2 dB, p < 0.0001) but relatively normal integrated backscatter from the media (-31.2 +/- 1.6 dB; p = NS versus normal aortas). The thickness of the media for both normal and atherosclerotic rabbits was approximately 300 microns. Histological characteristics of atherosclerotic aortas confirmed the presence of substantial intimal thickening, with prominent foam cell and lipid infiltration abutting a more normal medial layer.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Acoustic microscopy of human metaphase chromosomes.

Acoustic micrographs of human metaphase chromosomes have been recorded with wavelengths as short as 470 nm using liquid argon near 85 K as the acoustic wave coupling medium. Chromosomes prepared by trypsin-Giemsa staining exhibit acoustic banding patterns similar to the G-bands seen in optical images. Unstained chromosomes exhibit acoustic markings that do not correspond to traditional banding patterns. The observed acoustic contrast may arise from spatial variations in chromosomal mechanical properties, or from thickness variations in the fixed chromosomes.

Azure Stains↗

Acoustic inhomogeneity of carotid arterial plaques determined by GHz frequency range acoustic microscopy.

Characterization of the structure and composition of carotid arterial plaques is important to predict the risk of embolic and thrombotic events. In clinical echography, echolucent lesions seem to be more associated with stroke than echogenic lesions, and a low grey-scale median is associated with a fivefold increase in the incidence of silent infarcts. In the present study, five fixed human carotid atherosclerotic lesions were observed by GHz-range scanning acoustic microscopy (SAM). The atherosclerotic lesions were characterized by either thickened fibrosis with dense collagen fibers or lipid accumulation with sparse collagen network by optical microscopy. SAM revealed that the fibrosis was classified into type I and III collagen by attenuation of ultrasound (US) and that the sound field of lipid accumulation lesions became inhomogeneous. The results would provide the scientific basis for the images of vulnerable plaques being produced in diagnostic US.

Acoustics↗

Cell wall appositions and plant disease resistance: Acoustic microscopy of papillae that block fungal ingress.

Plant cells react to localized stress by forming wall appositions outside their protoplasts on the inner surface of their cellulose walls. For many years it has been inferred that appositions elicited by encroaching fungi, termed "papillae," may subsequently also deter them and thus represent a disease-resistance mechanism. Recently, it has been shown that preformed, oversized papillae, experimentally produced in coleoptile cells of compatible barley, Hordeum vulgare, can completely prevent direct entry of Erysiphe graminis f. sp. hordei that ordinarily penetrates and causes disease. To discover how these papillae may function, acoustic microscopy was used to contrast their in vivo elastic properties with those of ineffective normal papillae and contiguous cell wall. Raster and line scans showed intense acoustic activity at sites of preformed papillae; scans in selected focal planes identified this activity with the papillae, not with subtending cell wall. Minimal acoustic activity was found in normal papillae. It is suggested that some wall appositions could serve in disease resistance as viscoelastic barriers to mechanical forces exerted by the special penetration structures of advancing pathogenic fungi.

Journal Article↗

Relationship between myocardial tissue density measured by microgravimetry and sound speed measured by acoustic microscopy.

If myocardial tissue can be assumed to be fluid-like, myocardial tissue elasticity can be estimated by the sound speed of tissue based on the equation K = rho(c)2, where K is the elastic bulk modulus, rho is density, and c is the sound speed of tissue. However, little data exist regarding the relationship between the sound speed of tissue and tissue density. The purpose of the present study was to evaluate the relationship between the sound speed of tissue and tissue density of various diseased myocardia. Myocardial tissue specimens at autopsy were obtained from 10 control patients without cardiovascular disease, 8 patients with pressure overload left ventricular hypertrophy (POLVH), and 8 patients with cardiac amyloidosis (AMD). Myocardial tissue sound speed was measured using a scanning acoustic microscope operating in the frequency of 450 MHz, and tissue density was measured by microgravimetry. The sound speed in POLVH (1639 +/- 17 m/s) was higher and that in AMD (1565 +/- 11 m/s) was lower than that in control patients (1615 +/- 15 m/s) (p < 0.001) at the temperature of 20-22 degrees C. The density in POLVH (1.087 +/- 0.004 g/cm3) was higher and that in AMD (1.072 +/- 0.003 g/cm3) was lower than that in control patients (1.082 +/- 0.003 g/cm3) (p < 0.001). Tissue density correlated with sound speed in all three groups (r = 0.96, p < 0.001). Therefore, myocardial tissue sound speed data obtained by acoustic microscopy enabled us to evaluate tissue elasticity without measuring tissue density directly.

Acoustics↗

Combined surface-focused acoustic microscopy in transmission and scanning ultrasonic holography.

Employment of ultrasound techniques in nondestructive testing may require identification of the acoustic modes contributing to imaging. Such identification can be achieved, with some restrictions, by time-of-flight analysis. Another approach is acoustic holography that reveals the propagation properties of any selected mode. In anisotropic media, the propagation features are distinct and allow for a reliable classification of the selected mode. Both techniques were applied for classification of bonded, disbonded, and weakly bonded areas in directly bonded semiconductor wafers.

Equipment Design↗

Tension modulates cell surface motility: A scanning acoustic microscopy study.

The subtraction of subsequent scanning acoustic microscope images (SubSAM) of living cells taken in distinct time intervals reveals subcellular motility domains that are dependent on metabolic energy and correspond to cell surface deformations like protrusions, ruffling, and microblebs. This motility can be quantitated by determining the changes of the grey levels vs. time. Tension has been postulated as a global parameter in the control of cell shape and cell surface motility [Albrecht-Bühler 1987: Cell Motil. Cytoskeleton 7:54-67; Bereiter-Hahn et al., 1995: Biochem. Cell Biol. 73:337-348; Sheetz and Dai, 1996: Trends Cell Biol. 6:85-89]. For direct evaluation, the activity of the motility domains was measured while applying external tension (stretching) or internal tension (contraction induced by nocodazole) and by relaxation due to desintegration of the actin-cytoskeleton using low concentrations of cytochalasin D (0.5 microg/ml). Elevated tension, regardless of how it is generated, externally or internally, whether directed or isotropic, lowers cell surface motility. In contrast, the relaxation of the cell cortex by cytochalasin D increases cell surface motility. Thus, a direct relationship between tension at the cell surface and surface motility was established as has been suggested by Sheetz and Dai [1996: Trends Cell Biol. 6:85-89]

Adenosine Triphosphate↗

Possible temperature effects computed for acoustic microscopy used for living cells.

Imaging of living cells or tissues at a microscopic resolution, where GHz frequencies are used, provides a foundation for many new biological applications. The possible temperature increase causing a destructive influence on the living cells should be then avoided. However, there is no information on possible local temperature increases at these very high frequencies where, due to strongly focused ultrasonic beams, nonlinear propagation effects occur. Acoustic parameters of living cells were assumed to be close to those of water; therefore, the power density of heat sources in a water medium was determined as a basic quantity. Hence, the numerical solution of temperature distributions at the frequency of 1 GHz was computed for high and low powers generated by the transducer equal to 0.32 W and 0.002 W. In the first case, typical nonlinear propagation effects were demonstrated and, in the second one, propagation was almost linear. The focal temperature increase obtained in water equaled 14 degrees C for the highest possible theoretical repetition frequency of fr = 10 MHz and for the thermal insulation at the sapphire lens-water boundary. Simultaneously, the scanning velocity of the tested object was assumed to be incomparably low in respect to the acoustic beam velocity. The maximum temperature increase in water occurred exactly at this boundary, being equal there to 20 degrees C. It was shown that, first of all, the very high absorption of water was significant for the temperature distribution in the investigated region, suppressing the focal temperature peaks. Because the temperature increases are proportional to the repetition frequency, so for example, at its practical value of fr = 0.1 MHz, all temperature increases will be 100 times lower than listed above. For the low transducer power of 0.002 W, the corresponding temperature increases were about 140 times lower than those for the high power of 0.32 W. The presented solutions are devoted mainly to the reflection pulse mode; however, they can be also applied for the transmitting (continuous-wave) mode, as shown in an example. Pressure distributions were computed for the acoustic field of the microscope for the first and higher harmonics. Hence, at the frequency of 1 GHz, the effective focal radius in water measured as the -6-dB amplitude pressure drop was found to be 1,1 microm, and 0.7 microm for the second harmonic, independently of the assumed transducer power. So the width of the beam, scanning the living cells in the focal region, was equal to 2.2 microm at the fundamental frequency of 1 GHz.

Acoustics↗

A comparative study of the application of scanning acoustic microscopy and confocal laser scanning microscopy to the structural assessment of human bones.

The Scanning Acoustic Microscope (SAM) is used to image mechanical structure and to measure the sound velocity in micrometers of in vitro samples from human bone. A similar principle is used in confocal laser scanning microscopy. The same sample is imaged by both methods and the results compared. We concentrated on the SAM, the function of which is described in greater detail. We demonstrate that the confocal scanning principle is applied in both microscopes, although information about structure differs, depending on the method used.

Acoustics↗

Autocorrelation length of normal and myopathic human myocardium measured by acoustic microscopy: implications for clinical ultrasonic tissue characterization.

The pattern of acoustic speckle present in ultrasonic images of tissue may be affected by pathological changes in the tissue. To investigate the possibility for such an effect in dilated cardiomyopathy, we examined acoustic micrographs of frozen sections of normal (autopsy) and myopathic (transplant recipient) human myocardium. The resulting images were digitized, and the major axis autocorrelation length calculated for each image. For the normal specimens (n = 75, four patients), the mean autocorrelation length was 37 +/- 18 microns, with the maximum value of 92 microns. The myopathic specimens (n = 64, four patients) had a mean of 52 +/- 19 microns, with a maximum of 100 microns. We conclude that the changes in structure between normal and myopathic myocardium are significant (p less than 0.001), but on so small a scale in relation to the wave length of clinical ultrasound that there will only be changes in the intensity of the backscattered signal, not the pattern of speckle.

Acoustics↗

The use of acoustic microscopy for biological tissue characterization.

A system of transmission raster acoustic microscope with an ultrasound frequency of 450 MHz has been designed to investigate biological tissues and comparative analysis of their optical and acoustic images. The possibility of obtaining the contrast acoustic images of nonfixed, nonstained biological tissues and viscoelasticity measurements in microscale was demonstrated.

Animals↗