Faculty Bibliography

We have demonstrated the utility of ultrasound backscatter microscopy for targeted intraparenchymal injections into embryonic day (E) 13.5 mouse embryos. This system has been used to test the degree of commitment present in neural progenitors from the embryonic ventral telencephalon and mid-hindbrain region. Many E13.5 ventral telencephalic progenitors were observed to integrate and adopt local phenotypes following heterotopic transplantation into telencephalic or mid-hindbrain targets, whereas mid-hindbrain cells of the same stage were unable to integrate and change fate in the telencephalon. In contrast, many mid-hindbrain cells from an earlier developmental stage (E10.5) were capable of integrating and adopting a forebrain phenotype after grafting into the telencephalon, suggesting that mouse mid-hindbrain progenitors become restricted in their developmental potential between E10.5 and E13.5

The authors have modified a 40-50 MHz ultrasound imaging system to allow image-guided injections into targeted brain regions of living mouse embryos. The injection technique is described, and example injections are shown. The significance of this research in basic biological science and the potential impact on future attempts at in utero therapy are described

Extensive genetic information and the recent introduction of transgenic techniques for genetic manipulation have made the mouse the accepted model for normal cardiac development and congenital heart disease. An impediment to progress in this area has been the lack of noninvasive technologies to assess embryonic cardiovascular structure and function. We have developed a high frequency (40-50 MHz) ultrasound imaging and Doppler system enabling in vivo analysis of early embryonic cardiovascular development in the mouse. Simultaneous real-time image and Doppler data can be obtained noninvasively allowing, for the first time, measurements to be made of inflow and outflow characteristics of the mouse embryo heart and blood flow in the umbilical circulation. High resolution (measured lateral resolution = 60-100 ?m) images are used to position the sample volume of the Doppler transducers. The 40 MHz CW Doppler system utilizes a pair of air backed LiNbO3 transducers mounted on SMB connectors, and has been calibrated with a string phantom from 10 to 300 mm/s, velocities relevant to embryonic cardiovascular blood waveforms. Biphasic inflow waveforms were measured in normal 10 day mouse embryos (approximately equivalent to 4 weeks human) with peak blood velocities close to 200 mm/s. Outflow velocities in the same embryos were close to 70 mm/s while umbilical artery velocities were approximately 20 mm/s. Future studies will compare normal velocity data to measurements of mutant mice with specific defects in heart and placenta

BACKGROUND AND DESIGN: Ultrasound imaging, while initially developed to visualize internal organs, is now being applied to image the skin. In this preliminary study, we used a high-frequency, 40-MHz ultrasound imaging system to provide high-resolution images in psoriasis and examined the relationship between clinical and ultrasound ratings in plaque-type psoriasis. The ultrasound image of a psoriatic plaque demonstrates a superficial echogenic band (band A), followed by a nonchogenic band (band B), and a deeper echogenic band (band C). RESULTS: In psoriatic plaques (N = 145), the severity of the psoriasis as assessed according to the degree of scaling, erythema, and thickness (SET score) correlated best with the width of band B (P < .001, r = 0.86) and less well with the width of bands A (P < .001, r = 0.59) and C (P < .001, r = 0.44). For the treated psoriatic plaques (n = 64), for which paired readings were available before and after therapy, changes in the SET scores correlated best with the change in the width of band B (P < .001, r = 0.96) and less well with the change in the width of bands A (P < .001, r = 0.61) and C (P < .001, r = 0.45). Ultrasound analyses and clinical evaluation were performed by independent raters. CONCLUSIONS: The data suggest that high-frequency ultrasound imaging may prove to be a noninvasive technique that can be used as an adjunct to the clinical evaluation of the lesional severity of psoriatic plaques

BACKGROUND. Ultrasound imaging systems operating close to 20 MHz in frequency have been used to image skin tumors. Ultrasound imaging at 20 MHz has been used to determine the boundaries of basal cell carcinomas (BCCs). An inherent shortcoming of imaging systems operating at these frequencies is their limited resolution. OBJECTIVE. We investigated whether 40-MHz ultrasound imaging could provide higher resolution compared with the lower frequency systems and thus be a superior, noninvasive method of assessing the boundaries of BCCs. METHODS. Nine BCCs from six individuals were examined clinically and ultrasonographically, and then biopsied to confirm diagnosis. The depth of BCCs measured on histological sections was compared with the corresponding value obtained using ultrasound. For this study we required a nonsurgical, nondestructive means of treating BCCs that would allow repeated ultrasound imaging, and therefore topical 5-flurouracil (5-FU) was chosen. Following 5-FU therapy a biopsy was obtained from the site of the treated BCC after ultrasound imaging had been performed. Clinical, ultrasonic and histopathologic evaluation of each BCC was carried out independently by different individuals. At the end of the study all the BCC sites were treated surgically be electrodesiccation and curettage or completely excised. RESULTS. High resolution ultrasound images of BCCs were obtained with agreement between histology and ultrasound findings in all none lesions prior to therapy and in eight of none lesions posttherapy. There was a significant correlation between the depth of BCCs measured histologically and using ultrasound (P = 0.0004, r = 0.92). CONCLUSIONS. This study suggests that 40-MHz ultrasound may provide an estimate of the boundary of a BCC in vivo. High frequency 40-MHz ultrasound imaging may be an adjunct to clinical and histologic evaluation but does not replace the need to obtain tissue for microscopic examination

The incidence and mortality rate of cutaneous melanoma continue to increase throughout the world, making the study of melanoma biology an important area of current research. While recent breakthroughs in transgenic mouse technology have led to promising mouse skin models of melanoma, there is presently no technique available for quantitatively studying subsurface melanoma progression, in vivo. We demonstrate the first application of an imaging method called ultrasound backscatter microscopy (UBM) for imaging early murine melanomas with spatial resolution of 30 microns axial and 60 microns lateral. Murine B16 F10 melanomas have been imaged from their earliest detection, over several days, until they are 2 to 5 mm in diameter. Melanoma dimensions measured by UBM were found to be in excellent agreement with those determined histopathologically on the excised tumours. The relative rms errors in UBM-determined melanoma height and width were found to be 8.7% and 4.2%, respectively. The mean rate of increase in tumour height of early murine melanoma was found to be 0.37 +/- 0.06 mm/day. Computer-generated volumetric renderings of melanomas have been produced from three-dimensional image data, allowing quantitative comparisons of tumour volumes to be made. Using a priori assumptions of ellipsoid tumour shape, the relative error in UBM-determined volume was shown to be less than 17%. These results should be of considerable interest to investigators studying melanoma biology using mouse skin models, and have implications in the use of high frequency ultrasound imaging for the clinical assessment of cutaneous melanoma

The assessment of cutaneous melanoma in the clinical setting is often difficult, and important features such as depth and width remain unknown until the pathology report is received. Access to prognostic features such as vertical height before excisional biopsy would offer a basis for guidance in defining surgical margins and early planning of treatment options. Recently developed high-frequency ultrasound imaging in the 40-to 60-MHz range is a noninvasive method that provides in vivo information about cutaneous lesions. Imaging at these frequencies provides high-resolution data within the range of the epidermis and dermis (3-4 mm in depth). Ten cutaneous melanomas and seven pigmented lesions were assessed in this fashion. Vertical height was documented and compared to histopathological findings. High-frequency ultrasound imaging determination of vertical height correlated well with the standard measurement of Breslow's thickness on histological sections only in midrange (1.0-3.0 mm) lesions. Inflammatory cells at the base of three melanomas provoked an overestimation of the depth measurement with ultrasonography. Thick keratin layers such as those found on the feet acted as a virtual block to the high-frequency scanner. The application of this new advance in noninvasive imaging technology to the clinical assessment of cutaneous melanoma provides interesting in vivo data but in its present state does not replace the need for the biopsy of pigmented lesions and histopathological diagnosis

The history of developmental and genetic analysis in the mouse has made it the model of choice for studying mammalian embryogenesis. Presently lacking is a simple technique for efficiently analyzing early mouse mutant phenotypes in utero. We demonstrate application of a real-time imaging method called ultrasound backscatter microscopy for visualizing mouse early embryonic neural tubes and hearts. This method was used to study live embryos in utero between 9.5 and 11.5 days of embryogenesis, with a spatial resolution close to 50 microns. Ultrasound backscatter microscope images of cultured embryos made it possible to visualize the heart chambers. This noninvasive imaging method was also used for analyzing a neural tube defect. The midhindbrain deletion associated with a null mutation of the Wnt-1 protooncogene was easily recognizable on ultrasound backscatter microscope images of 10.5- and 11.5-day embryos. Computer-generated volumetric renderings of the neural tube cavities were made from three-dimensional image data. This allowed a much clearer definition of the Wnt-1 mutant phenotype. These imaging techniques should be of considerable use in studying mouse development in utero

There is a growing interest in high resolution, subsurface imaging of cutaneous tissues using higher frequency ultrasound, and several commercial systems have been developed recently which operate at 20 MHz. Some of the possible applications of higher frequency skin imaging include tumour staging, boundary definition, and studies of the response of tumours to therapy, investigations of inflammatory skin conditions such as psoriasis and eczema, and basic studies of skin aging, sun damage and the effects of irritants. Investigation of these areas is quite new, and the role of ultrasound skin imaging is continuing to evolve. Lateral resolution in the 20 MHz imaging systems ranges from 200 to 300 microns, which limits imaging applications to cutaneous structures which are relatively large in size. In this paper, a real-time ultrasound backscatter microscope (UBM) for skin imaging is described which operates in the 40-100 MHz range, providing axial resolution between 17 and 30 microns and lateral resolution between 33 and 94 microns. This improvement in resolution over current skin ultrasound systems should prove useful in determining the margins of small skin lesions, and in obtaining more precise, in vivo skin thickness measurements to characterize nonmalignant skin disease. Example images of normal skin, seborrhoeic keratosis and malignant melanoma illustrate the imaging potential of this system

Two-dimensional (2-D) arrays have been proposed as a solution to the degradation in medical ultrasound image quality occurring as a result of asymmetric focusing properties of linear phased array transducers. The 2-D phased transducer array is also capable of electronically steering the symmetrically focused ultrasound beam throughout a three-dimensional volume. In a companion paper the potential of 2-D transducer arrays for medical imaging has been investigated using simulated B-scan images. In this paper, the advantages of 2-D over linear transducer arrays is demonstrated by simulating images of spherical cysts embedded in a large scattering volume. The large elevation beamwidth in the nearfield of a 5 MHz linear phased transducer array results in a severe reduction in the image contrast measured between a 4 mm diameter cyst and the surrounding scattering media. By employing a 2-D array with symmetric focusing, the contrast between the cyst and surrounding scatterers is significantly improved. The use of additional elements in the elevation direction of a linear array is also investigated. In this case the additional elements are included only to focus, but not to steer the ultrasound beam. Using the contrast characteristics of a 4 mm diameter cyst, it is shown that relatively few elevation elements are required to significantly improve the nearfield imaging capability of the linear array