Yoshifumi Saijo

Ultrasonic tissue characterization of infarcted myocardium by scanning acoustic microscopy

The purpose of this study was to ultrasonically characterize infarcted human myocardial tissue at the microscopic level by scanning acoustic microscopy. Infarcted myocardial specimens from ten cases with acute myocardial infarction were studied. Specimens were formalin fixed, paraffin embedded and sectioned to 10-micron thickness. A specially developed scanning acoustic microscope system, operating in the 100- to 200-MHz ultrasound frequency range, was used for the measurements. The values of the attenuation constant were 0.94 +/- 0.04 dB/mm/MHz in normal myocardium, 0.71 +/- 0.12 dB/mm/MHz in degenerated myocardium, 0.88 +/- 0.47 dB/mm/MHz in granulation tissue and 1.75 +/- 0.11 dB/mm/MHz in fibrosis. The values of sound speed were 1620.2 +/- 8.2 m/s in normal myocardium, 1572.4 +/- 10.6 m/s in degenerated myocardium, 1590.2 +/- 32.5 m/s in granulation tissue and 1690.3 +/- 9.1 m/s in fibrosis. The ultrasonic properties of the diseased myocardium at the microscopic level will provide important information for ultrasonic tissue characterization at the macroscopic level.

The ultrasonic properties of gastric cancer tissues obtained with a scanning acoustic microscope system

A newly developed scanning acoustic microscope (SAM) system operating in the frequency range of 100-200 MHz has been employed to measure the attenuation and the sound speed of formalin-fixed specimens of five different types of gastric cancer. Signet-ring cell carcinoma specimens exhibit attenuation constant and sound speed values significantly lower than other types of gastric cancer tissues. Tubular adenocarcinoma specimens exhibit a trend toward higher attenuation and sound speed values as the cell type became differentiated. Our measurements and observations suggest that the ultrasonic properties are influenced by cellular arrangement, intercellular junction and intracellular chemical components.

ACOUSTIC PROPERTIES OF ATHEROSCLEROSIS OF HUMAN AORTA OBTAINED WITH HIGH-FREQUENCY ULTRASOUND

The ultrasonic properties of the tissue elements in the aorta were measured using a scanning acoustic microscope (SAM). Twelve autopsied aortas were formalin-fixed, frozen and sectioned at 10 microm thickness and mounted on glass slides for SAM investigation. A specially developed SAM system operating in the frequency range of 100-200 MHz was employed, and color-coded images of the two-dimensional (2-D) distributions of attenuation and sound speed were displayed. The region-of-interest (ROI) for attenuation and sound speed measurements was determined by comparison of optical and acoustic images. The average value of the slope of attenuation was 0.61 dB/mm/MHz and the sound speed was 1568 m/s in the normal intima; 2.5 dB/mm/MHz, 1760 m/s in the calcificated lesion; 1.7 dB/mm/MHz and 1677 m/s in the fibrosis; and 0.34 dB/mm/MHz, 1526 m/s in the fatty material, respectively. Acoustic microscopy provides the basic data for understanding the IVUS imaging of atherosclerosis, as well as on the pathological features of atherosclerosis.

Changes of articular cartilage after immobilization in a rat knee contracture model

The objective was to determine the changes of articular cartilage of the knee joint during immobilization in a rat model. The knee joints of adult male rats were immobilized at 150° of flexion using an internal fixator for 3 days, and 1, 2, 4, 8, and 16 weeks. The articular cartilage from the medial midcondylar region of the knee was obtained, divided into three areas (non‐contact area, transitional area, contact area), and in each area, a degree of degeneration was evaluated by gross observation, histomorphometric grading, and measurements of thickness and number of chondrocytes. Elasticity of the articular cartilage was estimated by measuring the sound speed with use of scanning acoustic microscopy. Degeneration of the articular cartilage was mainly observed in the contact and transitional areas. Matrix staining intensity by safranin‐O and number of chondrocytes were decreased in these two areas. The thickness of the articular cartilage in the non‐contact and contact areas was unchanged, but it was increased in the transitional area. Decrease in sound speed was observed in the transitional area of both the femoral and tibial cartilage, indicating the softening of the articular cartilage. The changes of articular cartilage became obvious as early as 1 week after immobilization. These changes may be due to a lack of mechanical stress or a lack of joint fluid circulation during immobilization. Although we do not know the reversibility of these changes of articular cartilage, early mobilization is preferable to avoid these cartilage changes.

Ultrasonic Tissue Characterization of Atherosclerosis by a Speed-of-Sound Microscanning System

We have been developing a scanning acoustic microscope (SAM) system for medicine and biology featuring quantitative measurement of ultrasonic parameters of soft tissues. In the present study, we propose a new concept sound speed microscopy that can measure the thickness and speed of sound in the tissue using fast Fourier transform of a single pulsed wave instead of burst waves used in conventional SAM systems. Two coronary arteries were frozen and sectioned approximately 10 mum in thickness. They were mounted on glass slides without cover slips. The scanning time of a frame with 300 X 300 pixels was 90 s and two- dimensional distribution of speed of sound was obtained. The speed of sound was 1680 plusmn 30 m/s in the thickened intima with collagen fiber, 1520 plusmn 8 m/s in the lipid deposition underlying the fibrous cap, and 1810 plusmn 25 m/s in a calcified lesion in the intima. These basic measurements will help in the understanding of echo intensity and pattern in intravascular ultrasound images.

Fundamental study of ultrasonic-measurement-integrated simulation of real blood flow in the aorta.

Acquisition of detailed information on the velocity and pressure fields of the blood flow is essential to achieve accurate diagnosis or treatment for serious circulatory diseases such as aortic aneurysms. A possible way to obtain such information is integration of numerical simulation and color Doppler ultrasonography in the framework of a flow observer. This methodology, namely, Ultrasonic-Measurement-Integrated (UMI) Simulation, consists of the following processes. At each time step of numerical simulation, the difference between the measurable output signal and the signal indicated by numerical simulation is evaluated. Feedback signals are generated from the difference, and numerical simulation is updated applying the feedback signal to compensate for the difference. This paper deals with a numerical study on the fundamental characteristics of UMI simulation using a simple two-dimensional model problem for the blood flow in an aorta with an aneurysm. The effect of the number of feedback points and the feedback formula are investigated systematically. It is revealed that the result of UMI simulation in the feedback domain rapidly converges to the standard solution, even with usually inevitable incorrect upstream boundary conditions. Finally, an example of UMI simulation with feedback from real color Doppler measurement also shows a good agreement with measurement.

Application of scanning acoustic microscopy for assessing stress distribution in atherosclerotic plaque.

AbstractScanning acoustic microscopy (SAM) was equipped to assess the acoustic properties of normal and atherosclerotic coronary arteries. The SAM image in the atherosclerotic lesion clearly demonstrated that the sound speed was higher than that in the normal intima, and that the variation of elasticity was found within the fibrous cap of the plaque. Youngs elastic modulus of each region was calculated and the finite element analysis was applied to derive the stress distribution in these arterial walls. In a case of normal coronary artery, the stress was dominant in the intima and the distribution was rather homogeneous and in a case of atherosclerosis, high stress was concentrated to the relatively soft lesion in the fibrous cap overlying lipid pool. SAM provides information on the physical properties, which cannot be obtained by the optical microscope. The results would help in understanding the pathological features of atherosclerosis.

Influence of tissue preparation on the high-frequency acoustic properties of normal kidney tissue

The influence of various tissue preparations on the acoustic properties of normal kidney tissue at high frequencies was investigated. Eight surgically excised normal kidney tissue specimens were classified into three groups: (i) fresh, frozen section, (ii) formalin-fixed, frozen section and (iii) formalin-fixed, paraffin section. Scanning acoustic microscopy operating in the frequency range of 100-200 MHz was used to display the two-dimensional distribution of attenuation constant and sound speed. Our results indicate that (i) there is no significant variation in both acoustic parameters between the three tissue groups, (ii) fixation by 10% formalin produces no significant change in the acoustic parameters, (iii) in fat-free tissue regions, the acoustic parameters are independent of preparation method and (iv) frozen sections must be used to assess the acoustic parameters in fat-rich tissues.

Visualization of human umbilical vein endothelial cells by acoustic microscopy

The morphology and acoustic properties of the human umbilical vein endothelial cells (HUVECs) were evaluated using a scanning acoustic microscope system. HUVECs were cultured for 4 days and exposed to the endotoxin for 4 h. The frequency of the scanning acoustic microscope was variable between 100 and 210 MHz. By changing the measuring frequency, ultrasonic amplitude and phase were measured and the quantitative value of attenuation was calculated. Before and after endotoxin stimuli, HUVECs were observed by scanning acoustic microscopy and the attenuation was measured. The acoustic images were successfully obtained to identify the outer shape of the HUVEC and the location of the nucleus in the cell. The attenuation of the nucleus is higher than that of the cytoplasm. The attenuation of the cytoplasm was increased and became inhomogeneous after endotoxin exposure. This finding would be related to the change of F-actin filaments, which is the main component of the cytoskeleton. Scanning acoustic microscopy is useful for assessing the cellular viscoelastic properties since it can detect both the morphological and acoustic changes without contacting the cellular surface.

Increased Elasticity of Capsule After Immobilization in a Rat Knee Experimental Model Assessed by Scanning Acoustic Microscopy

Objectives: The mechanical property of immobilized joints is not well understood. The present study was designed to investigate the tissue elasticity of the anterior and posterior synovial membrane (SM) in a rat immobilized knee model using scanning acoustic microscopy (SAM). Moreover, the structural characteristics of the SM after immobilization were examined by transmission electron microscopy (TEM). Methods: Thirty rats had their knee joints immobilized with a plate and metal screws. The rats were fixed at 1, 2, 4, 8 and 16 weeks after surgery and the knee joints were sectioned sagittally for SAM. Selected specimens were processed for TEM. A new concept SAM using a single pulsed wave instead of continuous waves was applied to measure the sound speed of the anterior and posterior SM, comparing it with the corresponding light microscopic images. Results: The sound speed of the posterior SM increased significantly in the 8- and 16-week experimental group compared with that in the control group. The sound speed of the anterior SM showed no statistical difference between the experimental and the control groups at any period of immobilization. The posterior SM of the experimental group was different from that of the control group in the ultrastructural characteristics of extracellular matrices. Conclusions: Our data suggest that the increased elasticity and structural changes of the posterior SM are one of the main causes of limited extension after a long period of immobilization in flexion using SAM, which is a powerful tool for evaluating the elasticity of targeted tissues.