Yi Sing Hsiao

Two-Bubble Acoustic Tweezing Cytometry for Biomechanical Probing and Stimulation of Cells

The study of mechanotransduction relies on tools that are capable of applying mechanical forces to elicit and assess cellular responses. Here we report a new (to our knowledge) technique, called two-bubble acoustic tweezing cytometry (TB-ATC), for generating spatiotemporally controlled subcellular mechanical forces on live cells by acoustic actuation of paired microbubbles targeted to the cell adhesion receptor integrin. By measuring the ultrasound-induced activities of cell-bound microbubbles and the actin cytoskeleton contractile force responses, we determine that TB-ATC elicits mechanoresponsive cellular changes via cyclic, paired displacements of integrin-bound microbubbles driven by the attractive secondary acoustic radiation force (sARF) between the bubbles in an ultrasound field. We demonstrate the feasibility of dual-mode TB-ATC for both subcellular probing and mechanical stimulation. By exploiting the robust and unique interaction of ultrasound with microbubbles, TB-ATC provides distinct advantages for experimentation and quantification of applied forces and cellular responses for biomechanical probing and stimulation of cells.

Dual-wavelength photoacoustic technique for monitoring tissue status during thermal treatments.

Abstract. Photoacoustic (PA) techniques have been exploited for monitoring thermal treatments. However, PA signals depend not only on tissue temperature but also on tissue optical properties which indicate tissue status (e.g., native or coagulated). The changes in temperature and tissue status often occur simultaneously during thermal treatments, so both effects cause changes to PA signals. A new dual-wavelength PA technique to monitor tissue status independent of temperature is performed. By dividing the PA signal intensities obtained at two wavelengths at the same temperature, a ratio, which only depends on tissue optical properties, is obtained. Experiments were performed with two experimental groups, one with untreated tissue samples and the other with high-intensity focused ultrasound treated tissue samples including thermal coagulated lesion, using ex vivo porcine myocardium specimens to test the technique. The ratio of PA signal intensities obtained at 700 and 800 nm was constant for both groups from 25 to 43°C, but with distinct values for the two groups. Tissue alteration during thermal treatment was then studied using water bath heating of tissue samples from 35 to 60°C. We found that the ratio stayed constant before it exhibited a marked increase at around 55°C, indicating tissue changes at this temperature.

Characterization of the pancreas in vivo using EUS spectrum analysis with electronic array echoendoscopes

BACKGROUND Spectral analysis of the radiofrequency (RF) signals that underlie grayscale EUS images has been used to provide quantitative, objective information about tissue histology. OBJECTIVE Our purpose was to validate RF spectral analysis as a method to distinguish between chronic pancreatitis (CP) and pancreatic cancer (PC). DESIGN AND SETTING A prospective study of eligible patients was conducted to analyze the RF data obtained by using electronic array echoendoscopes. PATIENTS Pancreatic images were obtained by using electronic array echoendoscopes from 41 patients in a prospective study, including 15 patients with PC, 15 with CP, and 11 with a normal pancreas. MAIN OUTCOME MEASUREMENTS Midband fit, slope, intercept, correlation coefficient, and root mean square deviation from a linear regression of the calibrated power spectra were determined and compared among the groups. RESULTS Statistical analysis showed that significant differences were observable between groups for mean midband fit, intercept, and root mean square deviation (t test, P < .05). Discriminant analysis of these parameters was then performed to classify the data. For CP (n = 15) versus PC (n = 15), the same parameters provided 83% accuracy and an area under the curve of 0.83. LIMITATIONS Moderate sample size and spatial averaging inherent in the technique. CONCLUSIONS This study shows that mean spectral parameters of the backscattered signals obtained by using electronic array echoendoscopes can provide a noninvasive method to quantitatively discriminate between CP and PC.

TOMOGRAPHIC RECONSTRUCTION OF TISSUE PROPERTIES AND TEMPERATURE INCREASE FOR HIGH-INTENSITY FOCUSED ULTRASOUND APPLICATIONS

The acoustic and thermal properties as well as the temperature change within a tissue volume during high-intensity focused ultrasound ablation are critically important for treatment planning and monitoring. Described in this article is a tomographic reconstruction method used to determine the tissue properties and increase in temperature in a 3-D volume. On the basis of the iterative finite-element solution to the bioheat equation coupled with Tikhonov regularization techniques, our reconstruction algorithm solves the inverse problem of bioheat transfer and uses the time-dependent temperature measured on a tissue surface to obtain the acoustic absorption coefficient, thermal diffusivity and temperature increase within the subsurface volume. Numerical simulations were performed to validate the reconstruction algorithm. The method was initially conducted in ex vivo experiments in which time-dependent temperature on a tissue surface was measured using high-resolution, non-invasive infrared thermography.

Photoacoustic imaging: a potential new platform for assessment of bone health

The ultimate goal of this work is to develop a novel photoacoustic (QPA) platform for highly-sensitive and quantitative assessment of bone health. First, the feasibility to perform 3D photoacoustic imaging (PAI) of bone was investigated. Then another two techniques, including thermal photoacoustic measurement (TPAM) and photoacoustic spectral analysis (PASA), both being able to achieve quantitative results were investigated for bone characterization. TPAM, by evaluating the dependence of photoacoustic signal amplitude on the sample temperature, is sensitive to the chemical constituents in tissue and holds promise for assessment of bone mineral density (BMD). PASA characterizes micron size physical features in tissue, and has shown feasibility for objective assessment of bone microarchitecture (BMA). This integrated QPA platform can assess both bone mass and microstructure simultaneously without involving invasive biopsy or ionizing radiation. Since QPA is non-ionizing, non-invasive, and has sufficient penetration in both soft tissue and bone, it has unique advantages for clinical translation.

Bone assessment via thermal photoacoustic measurements

The feasibility of an innovative biomedical diagnostic technique, thermal photoacoustic (TPA) measurement, for nonionizing and non-invasive assessment of bone health is investigated. Unlike conventional photoacoustic PA methods which are mostly focused on the measurement of absolute signal intensity, TPA targets the change in PA signal intensity as a function of the sample temperature, i.e. the temperature dependent Grueneisen parameter which is closely relevant to the chemical and molecular properties in the sample. Based on the differentiation measurement, the results from TPA technique is less susceptible to the variations associated with sample and system, and could be quantified with improved accurately. Due to the fact that the PA signal intensity from organic components such as blood changes faster than that from non-organic mineral under the same modulation of temperature, TPA measurement is able to objectively evaluate bone mineral density (BMD) and its loss as a result of osteoporosis. In an experiment on well established rat models of bone loss and preservation, PA measurements of rat tibia bones were conducted over a temperature range from 370 C to 440 C. The slope of PA signal intensity verses temperature was quantified for each specimen. The comparison among three groups of specimens with different BMD shows that bones with lower BMD have higher slopes, demonstrating the potential of the proposed TPA technique in future clinical management of osteoporosis.

Lymph node characterization in vivo using endoscopic ultrasound spectrum analysis with electronic array echo endoscopes

Our purpose was to demonstrate the use of radiofrequency spectral analysis to distinguish between benign and malignant lymph nodes with data obtained using electronic array echo endoscopes, as we have done previously using mechanical echo endoscopes. In a prospective study, images were obtained from eight patients with benign-appearing lymph nodes and 11 with malignant lymph nodes, as verified by fine-needle aspiration. Midband fit, slope, intercept, correlation coefficient, and root-mean-square (RMS) deviation from a linear regression of the calibrated power spectra were determined and compared between the groups. Significant differences were observable for mean midband fit, intercept, and RMS deviation (t test P < 0.05). For benign (n = 16) vs. malignant (n = 12) lymph nodes, midband fit and RMS deviation provided classification with 89 % accuracy and area under receiver operating characteristic (ROC) curve of 0.95 based on linear discriminant analysis. We concluded that the mean spectral parameters of the backscattered signals from electronic array echo endoscopy can provide a noninvasive method to quantitatively discriminate between benign and malignant lymph nodes.

Infrared thermography for noninvasive real-time monitoring of HIFU ablation

Infrared imaging for spatiotemporal temperature measurements was explored in this study for non-contact monitoring of temperature increases generated by HIFU ablation. Using ex vivo cardiac tissue specimens, we investigated the correlations between the occurrence of events during HIFU ablation (e.g., lesion formation, cavity formation) and the 2D spatiotemporal temperature of the tissue surface measured during HIFU ablation from an infrared camera. An increase in the rate of temperature rise was observed when lesions formed at or slightly beneath the tissue surface. Spatial shifts in the maximum temperature location away from the HIFU focus were often observed with continuing HIFU exposure after lesion formation, suggesting tissue dehydration and cavitation formation during ablation with excessive heating.