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Evaluation of ultrasonic bubble detectors in vitro using calibrated microbubbles at selected velocities.

Two standard Doppler ultrasonic devices, currently used for detecting bubbles in vivo, have been evaluated and compared in vitro using carefully calibrated uniform micro-bubbles rising at terminal velocity through a static aqueous medium. Two unexpected findings were observed: (a) the focal length of the transducer apparently decreases for smaller bubble sizes, and (b) a significant horizontal convection current was produced by one of the instruments. When the medium was in motion, it was found that the sensitivity varied markedly with bubble velocity, varying from a minimum detectable diameter of 40 mum at 55 cm/sec to 170 mum at 20 cm/sec. These findings are discussed with regard to the limitations of the Doppler technique for monitoring gas emboli in vivo and as an early warning for decompression sickness in divers.

Decompression Sickness

Measurements of frequency spectra of transmission coefficients to ultrasound through trapped microbubbles.

Techniques which use hydrophobic polycarbonate thin sheets containing randomly spaced, fairly uniform small pores immersed in water to trap air bubbles have been found to be useful in biophysical experiments. The utilization of broadband polyvinylidene fluoride transducers in this work made it possible to measure a continuous frequency spectrum of the transmission coefficient of the trapped bubbles. The results of the measurements show: (1) the frequency response curve of the bubble ensemble is much broader than that of a single bubble predicted by theory; and (2) as the incident sound pressure at a micropore membrane increases from 110 to 660 Pa the resonance frequency of bubbles shifts to lower values by as much as 7%.

Air

Imaging microbubbles and tissues using a linear focussed scanner operating at 20 MHz: possible implications for the detection of cavitation thresholds.

B-Scan images have been used as a means of detecting the onset of cavitation in tissues. New echoes appearing during simultaneous sonication with therapeutic ultrasound have often been attributed to bubble formation. In this study temperature rises up to 15 degrees C W-1 cm2 in guinea pig leg (post-mortem) were found, which would itself increase the rate of growth of gas bubbles. It is also hypothesized that a temperature dependent attenuation coefficient predicted by Bamber and Hill may, at least in part, account for new echoes appearing in tissues. Possible artefactual echoes in the B-scan images arising from acoustically induced gas bubbles in the coupling medium are also discussed.

Animals

Contrast agents for diagnostic ultrasound: development and evaluation of polymer-coated microbubbles.

Although the concept of an ultrasound contrast agent dates from Gramiak's work in 1968 in which indocyanine green was injected into the ascending aorta and heart, no universally accepted contrast agent for ultrasound now exists. This is primarily due to problems with stability, size and/or toxicity of the agents which have been investigated. Development of an effective ultrasound contrast agent would be highly significant for the health care industry, since it would greatly expand the scope of ultrasound (a noninvasive and safe procedure) as a diagnostic technique. While encapsulated gas bubbles offer particular advantages in stability over hand-agitated systems, they frequently present problems with size. Capsules larger than 10 microns in diameter become entrapped in the capillary bed of the lung. This paper describes the use of ionotropic gelation of the naturally occurring polysaccharide, alginate, for microencapsulation of air. Two procedures have been investigated. A novel jet head has been developed which allows co-extrusion of a solution of sodium alginate and air to produce nascent microencapsulated air bubbles which fall into a hardening solution of calcium ions. A second method employs ultrasound to introduce cavitation-induced bubbles into the alginate before capsule formation by spraying. Power spectra of these preparations demonstrate echogenicity (that is strong scatter of the incident ultrasound wave back to the emitting transducer, which also acts as a receiver), with resonant peaks that are a function of capsule size and wall characteristics.

Alginates

Echocardiographic contrast agents: effect of microbubbles and carrier solutions on left ventricular contractility.

Recently, there has been a resurgence of interest in the use of contrast-enhanced echocardiography as a means of noninvasively assessing myocardial perfusion. However, if injections of echocardiographic contrast agents are to be used for this purpose it is essential that they are not intrinsically toxic to the heart. In this study, the left ventricular end-systolic wall stress-rate-corrected velocity of fiber shortening relation, a load independent index of contractility, was studied in nine dogs. Two-dimensional and targeted M-mode echocardiographic as well as central aortic pressure tracings were made during echocardiographically gated, pressure- and volume-controlled aortic root injections of nonsonicated and sonicated Renografin-76, saline and dextrose 70% (n = 6), and sonicated and hand-agitated Renografin-76/saline mixture (n = 5). Two of nine dogs received all agents. Off-line computer videodensitometric analysis documented myocardial perfusion. In all cases, data were obtained at control and 5 and 15 seconds after injection. Additional data were collected at 25 seconds after injection for the Renografin-76/saline mixture. Alterations in contractility were measured relative to control as changes in rate-corrected velocity of fiber shortening after afterload (measured as end-systolic wall stress) was eliminated as a confounding variable. Under no condition did saline or Renografin-76 cause alterations in left ventricular contractility. Nonsonicated and sonicated dextrose 70% increased left ventricular contractility at 15 seconds but not at 5 seconds after injection. Hand-agitated Renografin-76/saline mixture induced a negative inotropic effect at 5 and 15 seconds after injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

In vivo microbubble detection in decompression sickness using a second harmonic resonant bubble detector.

A resonant bubble detection method based on a second harmonic technique has been used to monitor the femoral vascular system of dogs subjected to rapid decompression. For this study, the detector consisted of two acoustic transducers mounted at right angles to each other that were packaged in a perivascular cuff configuration. This detector responds selectively only to bubbles near resonant size (4.2 mum in diameter); solid particles and large bubbles produce no response. The detector was used to monitor a total of 15 dogs. Eleven dogs were subjected to a series of simulated underwater dives until acute symptoms of decompression sickness occurred; 4 dogs served as controls. In the dived group, either the femoral vein or the femoral artery was monitored. Resonant bubbles were observed in the femoral veins of all 6 dogs monitored at this location. During arterial monitoring, most dogs showed no response, but an occasional weak response was observed in 2 of the dogs. No resonant bubbles were detected in the femoral artery or the femoral vein in any of the controls. The data suggest that this bubble detection method is feasible for in vivo use. Furthermore, 4 mum diameter bubbles are much more prevalent in the veins of dogs suffering from decompression sickness than they are in dog arteries, presumably because they are filtered out effectively by the pulmonary circulation. Modifications of this method are discussed to enhance its accuracy and applicability for quantifying bubble size, location, and number.

Animals

The acoustic filter: an ultrasonic blood filter for the heart-lung machine.

Cardiopulmonary bypass-associated encephalopathy is thought to be due in part to continuous microembolization of the brain with gas microbubbles more than 40 microns in diameter during bypass. Current barrier filter technology cannot effectively remove such small microbubbles in fragile fluids such as blood. The design concepts for a new nonbarrier ultrasound-based fluid filtration system (an "acoustic filter") capable of filtering small microbubbles from blood are presented. The acoustic filter uses a field of high-intensity ultrasound to push microbubbles down an acoustic gradient, where they can be collected and removed. To test the filtration efficiency of the system, a Doppler ultrasound bubble detector was built. By monitoring the prefilter and postfilter Doppler signal an assessment of filtration efficiency was made. A suspension of stable albumin-encapsulated microbubbles (4 to 32 microns) were used as a model of the microbubble contaminants that might be found in the arterial return line of the heart-lung machine. Inactivated, the acoustic filter neither removed nor added microbubbles to the fluid. Activated, the acoustic filter provided total or near-total clearing of microbubbles. We conclude that the acoustic filter can remove microbubbles from a cardiopulmonary bypass-like apparatus.

Acoustics

The source of ultrasound contrast effect.

Evidence that microbubbles are the main sources of ultrasound contrast in injected solutions has been largely indirect. To investigate this directly, we examined freshly agitated indocyanine green, freshly agitated water, commercially prepared precision microbubbles (diameter 75 +/- 25 mu) in gelatin, carbonated water, "degassed" indocyanine green solution, and "degassed" water in one or more of four different assay systems. Only fluids with microbubbles produced ultrasound contrast. Injected contrast material rose in a water bath at a rate that identified it as being caused by microbubbles. Indocyanine green and gelatin surface tensions were measured and found to be low (43 dynes/cm2), thus explaining their tendency to stabilize the microbubbles that cause ultrasound contrast effect when injected and to hold foam after agitation. The force of hand injections (force similar to that used clinically through catheters and 19-gauge or 23-gauge needles) was below the force needed to cause cavitation or ultrasound contrast effect. Microbubble content could be quantified by the decrease in amplitude of the echo from a structure distant to the microbubbles. We conclude that that the ultrasound contrast effect seen in peripherally injected fluids is caused by microbubbles present in the injectant. The contrast is not due to cavitation at needle tips, and it can be quantified over a limited range. Improved design for a peripheral contrast agent is suggest.

Animals

Sonicated echocardiographic contrast agents: reproducibility studies.

This article describes the production, analysis, and reproducibility of forming microbubbles for contrast ultrasound imaging. The sonication method used to generate microbubbles was tested by four independent observers, and a subsequent laser particle counter analysis of microbubble size and concentration determined the reproducibility of the method. The results indicated that the mean bubble size was 3.3 +/- 1.2 microns for the entire group, based on three trials of each of the four participants. The characteristics of the bubble size of the microbubbles between observers were assessed with a Poisson distribution with the reproducibility based on the sample mean for each observer's trials. Standardization and calibration of the laser particle counter was accomplished with commercially available latex spheres, sonicated albumin microspheres, and a Coulter counter analysis. Our results indicate that the sonication technique generates small microbubbles with a reproducible uniform size distribution. The method of microbubble production is reproducible and can be widely applied for use in contrast echocardiographic perfusion imaging of tissue in a variety of research and clinical studies.

Air