In vivo biomicroscopy with ultrasound 2

In the first article of this series it was shown that the use of inverse scattering theory to analyse ultrasound reflections could provide high resolution images of the acoustic impedance profile of the retina. Unlike the retina, most tissue structures of interest, like small tumours and arterial plaque deposits, are shielded from view by intervening layers of tissue of appreciable acoustic impedance and attenuation. By analysing a one-dimensional model for a plaque deposit on the wall of a carotid artery embedded in a 5 cm thick layer of tissue, we demonstrate that a relatively high quality image can be recovered when compensation for the attenuation of the intervening tissue is made. We observe that because of the dearth of low frequency power in the recovered signal of ultrasound transducers, it is important that the field of view imaged is not taken to be too large. We compare the exact iterative distorted wave Born approximation inverse scattering method with the approximate but computationally faster plane wave Born approximation method and find that they give images of comparable quality for this model.     

Ultrasound biomicroscopy measurement of skin thickness change induced by cosmetic treatment with ultrasound stimulation

Moisturizing creams and lotions are commonly used in daily life for beauty and treatment of different skin conditions such as dryness and wrinkling, and ultrasound stimulation has been used to enhance the delivery of ingredients into skin. However, there is a lack of convenient methods to study the effect of ultrasound stimulation on lotion absorption by skin in vivo. Ultrasound biomicroscopy was adopted as a viable tool in this study to investigate the effectiveness of ultrasound stimulation on the enhancement of lotion delivery into skin. The forearm skin of 10 male and 10 female young subjects was tested at three different sites, including two lotion treatment sites with (Ultrasound Equipment – UE ON) and without (UE OFF) ultrasound stimulation and a control site without any lotion treatment. 1 MHz ultrasound with a duty cycle of 1.7%, a spatial peak temporal peak pressure of 195 kPa and an average power of 0.43 W was used for the stimulation. The skin thickness before, immediately after (0 min), and 15 and 30 min after the treatment was measured by an ultrasound biomicroscopic system (55 MHz). It was found that the skin thickness significantly increased immediately after the lotion treatment for both UE ON (from 1.379 ± 0.187 mm to 1.466 ± 0.182 mm, p < 0.001) and UE OFF (from 1.396 ± 0.193 mm to 1.430 ± 0.194 mm, p < 0.001) groups. Further comparison between the two groups revealed that the skin thickness increase of UE ON group was significantly larger than that of UE OFF group (6.5 ± 2.4% vs. 2.5 ± 1.3%, p < 0.001). Furthermore, it was disclosed that the enhancement of lotion delivery by ultrasound stimulation was more effective for the female subjects than the male subjects (7.6 ± 2.3% vs. 5.4 ± 2.0% immediately after treatment, p = 0.017). In conclusion, this study demonstrated that ultrasound biomicroscopy was a feasible method for studying the effectiveness of lotion treatment in vivo, and ultrasound stimulation was effective to enhance the rate of lotion absorption into skin.     

In vivo imaging of blood flow in the mouse Achilles tendon using high-frequency ultrasound

Objective

Achilles tendinitis is a common clinical problem with many treatment modalities, including physical therapy, exercise and therapeutic ultrasound. However, evaluating the effects of current therapeutic modalities and studying the therapeutic mechanism(s) in vivo remains problematic. In this study, we attempted to observe the morphology and microcirculation changes in mouse Achilles tendons between pre- and post-treatment using high-frequency (25 MHz) ultrasound imaging. A secondary aim was to assess the potential of high-frequency ultrasound in exploring therapeutic mechanisms in small-animal models in vivo.

Methods

A collagenase-induced mouse model of Achilles tendinitis was adopted, and 5 min treatment of continuous-mode low-frequency (45 kHz) ultrasound with 47 mW/cm² maximum intensity and 16.3 cm² effective beam radiating area was applied. The B-mode images showed no focal hypoechoic regions in normal Achilles tendons either pre- or post-treatment. The Doppler power energy and blood flow rate were measured within the peritendinous space of the Achilles tendon.

Conclusion

An increase in the microcirculation was observed soon after the low-frequency ultrasound treatment, which was due to immediate induction of vascular dilatation. The results suggest that applying high-frequency Doppler imaging to small-animal models will be an invaluable aid in explorations of the therapeutic mechanism(s). Our future work includes using imaging to assess microcirculation changes in tendinitis between before and after treatment over a long time period, which is expected to yield useful physiological data for future human studies.     

In vivo measurement of attenuation

We propose a new technique for the in vivo measurement of attenuation. The method is conceptually simple and can also be easily implemented on a real-time ultrasound unit. The technique is suggested by an analysis of the propagation of a Gaussian pulse in a medium which has frequency-dependent attenuation as well as dispersion (frequency-dependent velocity). If the medium has a loss factor which can be described by H(f) = exp(-eta[f]pX), where f is the frequency and O less than or equal to p less than or equal to 2 (valid for tissues and other objects of interest), then the pulse retains its Gaussian shape, shifting only its centre frequency and bandwidth. This suggests that by measuring the mean frequency of the reflected rf waveform in a window which is moved in depth we can obtain an estimate of the attenuation. Here we describe a particular hardware implementation of this technique which we have completed (based on measurement of zero-crossings) and present some preliminary in vivo measurements.     

In vivo interactions of magnetic nanoparticles with the blood-brain barrier

Targeted drug delivery to the brain parenchyma, i.e., in brain tumor patients, by means of magnetically supported carrier delivery through the tight vascular endothelium of the blood-brain barrier is of critical biomedical importance. We were interested in delineating the first steps in successful brain drug delivery, which focuses on the interactions between magnetically guided yet freely blood circulating nanoparticles and the blood-brain barrier. We employed an in vivo model to quantitatively determine changes in cerebrovascular flow rate and volume during magnetically guided exposure of circulating nanoparticles.     

In vivo ultrahigh-resolution optical coherence tomography

Ultrahigh-resolution optical coherence tomography (OCT) by use of state of the art broad-bandwidth femtosecond laser technology is demonstrated and applied to in vivo subcellular imaging. Imaging is performed with a Kerr-lens mode-locked Ti:sapphire laser with double-chirped mirrors that emits sub-two-cycle pulses with bandwidths of up to 350 nm, centered at 800 nm. Longitudinal resolutions of ~1mum and transverse resolution of 3mum, with a 110-dB dynamic range, are achieved in biological tissue. To overcome depth-of-field limitations we perform zone focusing and image fusion to construct a tomogram with high transverse resolution throughout the image depth. To our knowledge this is the highest longitudinal resolution demonstrated to date for in vivo OCT imaging.     

In vivo dark-field reflection-mode photoacoustic microscopy

Reflection-mode photoacoustic microscopy with dark-field laser pulse illumination and high-numerical-aperture ultrasonic detection is designed and implemented in noninvasively imaged blood vessels in the skin in vivo. Dark-field optical illumination minimizes the interference caused by strong photoacoustic signals from superficial structures. A high-numerical-aperture acoustic lens provides high lateral resolution, 45-120 microm in this system. A broadband ultrasonic detection system provides high axial resolution, estimated to be approximately 15 microm. The optical illumination and ultrasonic detection are in a coaxial confocal configuration for optimal image quality. The system is capable of imaging optical-absorption contrast as deep as 3 mm in biological tissue.     

In vivo measurement of retinal physiology with high-speed ultrahigh-resolution optical coherence tomography

Noninvasive in vivo functional optical imaging of the intact retina is demonstrated by using high-speed, ultrahigh-resolution optical coherence tomography (OCT). Imaging was performed with 2.8 microm resolution at a rate of 24,000 axial scans per second. A white-light stimulus was applied to the dark-adapted rat retina, and the average reflectivities from different intraretinal layers were monitored as a function of time. A 10%-15% increase in the average amplitude reflectance of the photoreceptor outer segments was observed in response to the stimulus. The spatial distribution of the change in the OCT signal is consistent with an increase in backscatter from the photoreceptor outer segments. To our knowledge, this is the first in vivo demonstration of OCT functional imaging in the intact retina.     

In vitro and in vivo investigations of targeted chemotherapy with magnetic nanoparticles

Magnetic drug targeting is a local drug delivery system. Electromicroscopic pictures document the ferrofluid enrichment in the intracellular space in vitro. In vivo experiments were performed in VX2 tumor-bearing rabbits using magnetic nanoparticles bound to mitoxantrone. High-pressure liquid chromatography (HPLC) analyses after magnetic drug targeting showed an increasing concentration of the chemotherapeutic agent in the tumor region compared to regular systemic chemotherapy.     

In vivo NMR in pharmaceutical research.

In vivo NMR techniques are currently well established in pharmaceutical research and will likely become increasingly important in the future, as they procure noninvasively morphological, physiological, and biochemical information. The status of magnetic resonance imaging (MRI) and spectroscopy (MRS) in drug development is discussed on the basis of the characterization and evaluation of a rat model of ischemic stroke and the development and profiling of drugs for cerebral ischemia in this model. It can be concluded that MRI is well suited for drug screening (quantitative determination of lesion size), while dynamic MRI and MRS techniques provide relevant information on the mechanism of drug actions. The possibility to follow changes, pathological and therapeutic, in the same individual is important from two points of view. First, variations due to interindividual differences may be eliminated, increasing the statistical power of the results. Second, dose and/or time dependence of a drug can be explored in the same individual. As a result, the number of animals required for a study will be reduced, which from both ethical and economic aspects is highly desirable.     

In vivo measurement of basilar membrane stiffness

Basilar membrane stiffness measurements were made in the base of the gerbil cochlea. Basilar membrane stiffness was determined by contacting the basilar membrane with a stainless steel needle (tip diameter 25 microns) attached to a force transducer, putting the needle/transducer structure through a low-frequency sinusoidal excursion with amplitude 5 or 25 nm, and measuring the restoring force exerted on the needle by the basilar membrane at the applied frequency. Stiffness was calculated as the amplitude of the restoring force divided by the amplitude of the excursion. Stiffness was measured over a 24-microns range of static displacements of the basilar membrane and is presented as stiffness versus static displacement. In cochleas that were not damaged during surgery the stiffness versus displacement characteristic usually had the following features: (1) an initial stiffness plateau with average stiffness 0.6 N/m; (2) a second plateau or level off with average stiffness 9.1 N/m; and (3) an increase in stiffness beyond the second plateau that was consistent with the theoretical stiffness-vs-displacement function of a beam. These features were present both pre- and post-mortem.     

In vitro effects of ultrasound with different energies on the conduction properties of neural tissue

The effect of ultrasound at various energy levels on the conduction properties of neural tissue is explored in this in vitro study. Excised sciatic nerves from the bullfrog were used for experiments. The nerves were stimulated by 3.5 MHz continuous wave ultrasound at 1, 2, and 3 W for 5 min. The peak-to-peak amplitude of the electrically evoked compound action potential (CAP) and the conduction velocity (CV) were measured in the nerves before and during ultrasound stimulation. The CV of the nerves increased by 5-20% for ultrasound stimulations at 1-3 W. The CAP amplitude increased by 8% during stimulation with 1 W ultrasound, and progressively decreased for 2 and 3 W ultrasound. This indicates that the effect of lower energy ultrasound increases both the CV and the CAP amplitude and that the reduction in the CAP amplitude for higher energy ultrasound is associated largely with ultrasonic thermal effects.     

Reproducibility of human eye length measurement by ultrasound in vivo.

This article deals with the reproducibility of the ultrasound pulse echo method for the measurement of intra-ocular distances. It analyses individual kinds of errors and evaluates the minimum size of an intra-ocular pathological formation which still could be traced by the pulse-echo method.     

In vivo EDXRF scanning analysis of human nail

This paper presents the results of a new technique for in vivo energy dispersive X‐ray fluorescence (EDXRF) scan analysis, applied to human fingernails. The scan employs a specially designed EDXRF spectrometer, which allows a concentration profile of the elements detected in a human nail. In order to carry out this technique, a group of nail fragments taken from different people was analyzed. The elements S, Ca, Cu, Fe, Cu, Zn, and Pb were detected in most of the samples. A bidimensional (x, y) scan was also performed on a whole removed nail in which the 2D spatial distribution of the detected elements was observed. Significant differences in some of the detected elements were noted. Minimum time of average detection per element was determined, based on the EDXRF spectra of the nail fragment. The time required to obtain an in vivo element profile of a typical nail was thus determined, applying the same geometry and acquisition conditions for all cases. The dose that the person undergoing this type of EDXRF scan analysis would be exposed to was also determined. Exposure time does not exceed 15 s, and the calculated administered dose is in the surface nail region of 0.1 mGy/s. The results of this study demonstrate that it is possible to carry out an in vivo X‐ray fluorescence scan analysis. This information may be used in different fields of medicine, such as nutrition and toxicology, and in other areas that establish a correlation between the concentration of the detected elements and certain diseases. Nail and hair are known to be ‘accumulating tissues’ unlike bodily fluids. In some aspects, nail analysis can be equal to a blood test. Copyright © 2014 John Wiley & Sons, Ltd.     

In vivo echo-planar imaging of rat spinal cord

An integrated approach to echo-planar imaging of rat spinal cord in vivo with a small field of view (FOV) is presented. This protocol is based on a multishot interleaved echo-planar imaging (EPI) sequence and includes: 1) use of an inductively coupled implantable coil for improved signal-to-noise ratio (SNR); 2) three-dimensional (3D) automatic shimming of the magnetic field over the spinal cord; and 3) post-acquisition data processing using a multireference scan for minimizing image artifacts. Some of the practical issues in implementing this protocol are discussed. This imaging protocol will be useful in characterizing the spinal cord pathology using techniques that are otherwise time-consuming, such as diffusion tensor imaging.     

用于活体温度评估的非线性超声热应变模型

热疗在肿瘤消融、高血压治疗等方面有重要应用价值.对热疗过程进行温度监控有利于实施合理的治疗规划,同时可提高治疗的安全性、减少副作用.利用基于超声回波偏移的热应变测温理论进行二维温度估计是一种常用的超声测温方法.但该理论基于组织热膨胀及声速随温度线性变化的假设,适用的温度测量范围一般限于37℃～50℃.该文基于升温组织的...     

超声抗早孕

控制人口是全球进入２１世纪时所面临的重大问题，我国科研人员率先提出了超声抗早孕的新概念，几年来，从超声工程设备开发到对小鼠，香猪及猴子超声抗早孕的动物实验，做了大量系统的研究工作，所获的研究成果，为超声抗早孕的临床应用展示出令工鼓舞的广阔前景。     

Bioleaching with ultrasound

Application of bioleaching of metals in a large-scale operation is reviewed briefly. Continued technical innovation is vital for the wider utilisation of this extraction process within the mineral industries. Therefore, the use of power ultrasound in combination with bioleaching (referred to as sonobioleaching) has been studied and is shown to be beneficial in the recovery of nickel from lean grade ores. The role of ultrasound in improving the benefits and lowering the drawbacks of bioleaching to an acceptable level are described. A possible mechanism for improving and intensifying this process is hypothesised.