A Preliminary Study of Quantitative Ultrasound for Cancer-Risk Assessment of Thyroid Nodules.

Background: Gray-scale, B-mode ultrasound (US) imaging is part of the standard clinical procedure for evaluating thyroid nodules (TNs). It is limited by its instrument- and operator-dependence and inter-observer variability. In addition, the accepted high-risk B-mode US TN features are more specific for detecting classic papillary thyroid cancer rather than the follicular variant of papillary thyroid cancer or follicular thyroid cancer. Quantitative ultrasound (QUS) is a technique that can non-invasively assess properties of tissue microarchitecture by exploiting information contained in raw ultrasonic radiofrequency (RF) echo signals that is discarded in conventional B-mode imaging. QUS provides quantitative parameter-value estimates that are a function of the properties of US scatterers and microarchitecture of the tissue. The purpose of this preliminary study was to assess the performance of QUS parameters in evaluating benign and malignant thyroid nodules. Methods: Patients from the Thyroid Health Center at the Boston Medical Center were recruited to participate. B-mode and RF data were acquired and analyzed in 225 TNs (24 malignant and 201 benign) from 208 patients. These data were acquired either before (167 nodules) or after (58 nodules) subjects underwent fine-needle biopsy (FNB). The performance of a combination of QUS parameters (CQP) was assessed and compared with the performance of B-mode risk-stratification systems. Results: CQP produced an ROC AUC value of 0.857 ± 0.033 compared to a value of 0.887 ± 0.033 (p=0.327) for the American College of Radiology Thyroid Imaging, Reporting and Data System (ACR TI-RADS) and 0.880 ± 0.041 (p=0.367) for the American Thyroid Association (ATA) risk-stratification system. Furthermore, using a CQP threshold of 0.263 would further reduce the number of unnecessary FNBs in 44% of TNs without missing any malignant TNs. When CQP used in combination with ACR TI-RADS, a potential additional reduction of 49 to 66% in unnecessary FNBs was demonstrated. Conclusion: This preliminary study suggests that QUS may provide a method to classify TNs when used by itself or when combined with a conventional gray-scale US risk-stratification system and can potentially reduce the need to biopsy TNs.

High-frame-rate volume imaging using sparse-random-aperture compounding.

High-frame-rate volume imaging (HFR-VI) aims to provide high-quality images with high-temporal information. Despite its potential, HFR-VI translation into clinical applications has been challenging due to the high cost of the equipment required to drive matrix probes with a large number of elements. The goal of this study is to introduce and test sparse-random-aperture compounding (SRAC), a technique that allows use of matrix probes with an ultrasound system that has fewer channels while maintaining high frame rates. Four scanning methods were implemented with a 256-channel system using a 4-to-1 multiplexer and a 3 MHz matrix probe with 1024 elements. These methods used three types of waves, either single-diverging waves (SDW), multiplane-diverging waves (MDW) or wide beams (WB); and were driven using one to four SRAC. All methods were also implemented in a 1024-channel multisystem. The main-lobe-to-side-lobe ratio (MLSLR) and the contrast ratio (CR) were studied using a string phantom and a CIRS phantom, respectively. The results showed an increase in the MLSLR and CR as a function of the number of SRAC. The multisystem provided the best results for the MLSLR. However, four SRAC outperformed the multisystem with respect to CR. The method using SDW provided the highest frame rates (i.e. 1875 and 7500 Hz for four and one SRAC, respectively), however it provided the lowest image quality. The two methods using MDWs showed a good compromise between image quality and frame rate (i.e. 187 to 750 Hz for four and one SRAC). WB provided the best image quality at the expense of frame rate (i.e. 18 to 75 Hz for four and one SRAC). Our results suggest that SRAC in combination with the tested scanning methods can provide a low-channel count alternative for HFR-VI systems and allows a tunable tradeoff between image quality and frame rate guided by the desired application.

Quantifying skin photodamage with spatial frequency domain imaging: statistical results.

We investigated the change in optical properties and vascular parameters to characterize skin tissue from mild photodamage to actinic keratosis (AK) with comparison to a published photodamage scale. Multi-wavelength spatial frequency domain imaging (SFDI) measurements were performed on the dorsal forearms of 55 adult subjects with various amounts of photodamage. Dermatologists rated the levels of photodamage based upon the photographs in blinded fashion to allow comparison with SFDI data. For characterization of statistical data, we used artificial neural networks. Our results indicate that optical and vascular parameters can be used to quantify photodamage and can discriminate between the stages as low, medium, and high grades, with the best performance of ∼70%, ∼76% and 80% for characterization of low- medium- and high-grade lesions, respectively. Ultimately, clinicians can use this noninvasive approach for risk assessment and frequent monitoring of high-risk populations.

Regional changes in the elastic properties of myopic Guinea pig sclera.

Biomechanical changes in the sclera likely underlie the excessive eye elongation of axial myopia. We studied the biomechanical characteristics of myopic sclera at the microscopic level using scanning acoustic microscopy (SAM) with 7-µm in-plane resolution. Guinea pigs underwent form-deprivation (FD) in one eye from 4 to 12 days of age to induce myopia, and 12-µm-thick scleral cryosections were scanned using a custom-made SAM. Two-dimensional maps of the bulk modulus (K) and mass density (ρ) were derived from the SAM data using a frequency-domain approach. We assessed the effect on K and ρ exerted by: 1) level of induced myopia, 2) region (superior, inferior, nasal or temporal) and 3) eccentricity from the nerve using univariate and multivariate regression analyses. Induced myopia ranged between -3D and -9.3D (Mean intraocular difference of -6.2 ±â€¯1.7D, N = 11). K decreased by 0.036 GPa for every 1.0 D increase in induced myopia across vertical sections (p < 0.001). Among induced myopia right eyes, K values in the inherently more myopic superior region were 0.088 GPa less than the inferior region (p = 0.002) and K in the proximal nasal region containing the central axis were 0.10 GPa less than temporal K (p = 0.036). K also increased 0.12 GPa for every 1 mm increase in superior vertical distance (p < 0.001), an effect that was blunted after 1 week of FD. Overall, trends for ρ were less apparent than for K. ρ values increased by 20.7 mg/cm3 for every 1.00 D increase in induced myopia across horizontal sections (p < 0.001), and were greatest in the region containing the central posterior pole. ρ values in the inherently more myopic superior region were 13.1 mg/cm3 greater than that found in inferior regions among control eyes (p = 0.002), and increased by 11.2 mg/cm3 for every 1 mm increase in vertical distance (p = 0.001). This peripheral increase in ρ was blunted after 1 week of FD. Scleral material properties vary depending on the location in the sclera and the level of induced myopia. Bulk modulus was most reduced in the most myopic regions (both induced myopia and inherent regional myopia), and suggests that FD causes microscopic local decreases in sclera stiffness, while scleral mass density was most increased in the most myopic regions.

Cerebellar folding is initiated by mechanical constraints on a fluid-like layer without a cellular pre-pattern.

Models based in differential expansion of elastic material, axonal constraints, directed growth, or multi-phasic combinations have been proposed to explain brain folding. However, the cellular and physical processes present during folding have not been defined. We used the murine cerebellum to challenge folding models with in vivo data. We show that at folding initiation differential expansion is created by the outer layer of proliferating progenitors expanding faster than the core. However, the stiffness differential, compressive forces, and emergent thickness variations required by elastic material models are not present. We find that folding occurs without an obvious cellular pre-pattern, that the outer layer expansion is uniform and fluid-like, and that the cerebellum is under radial and circumferential constraints. Lastly, we find that a multi-phase model incorporating differential expansion of a fluid outer layer and radial and circumferential constraints approximates the in vivo shape evolution observed during initiation of cerebellar folding.

Autoregressive Signal Processing Applied to High-Frequency Acoustic Microscopy of Soft Tissues.

Quantitative acoustic microscopy (QAM) at frequencies exceeding 100 MHz has become an established imaging tool to depict acoustical and mechanical properties of soft biological tissues at microscopic resolutions. In this study, we investigate a novel autoregressive (AR) model to improve signal processing and parameter estimation and to test its applicability to QAM. The performance of the AR model for estimating acoustical parameters of soft tissues (i.e., acoustic impedance, speed of sound, and attenuation) was compared to the performance of the Hozumi model using simulated ultrasonic QAM signals and using experimentally measured signals from thin (i.e., 12 and ) sections of human lymph-node and pig-cornea tissue specimens. Results showed that the AR and Hozumi methods performed equally well (i.e., produced an estimation error of 0) in signals with low, linear attenuation in the tissue and high impedance contrast between the tissue and the coupling medium. However, the AR model outperformed the Hozumi model in estimation accuracy and stability (i.e., parameter error variation and number of outliers) in cases of 1) thin tissue-sample thickness and high tissue-sample speed of sound, 2) small impedance contrast between the tissue sample and the coupling medium, 3) high attenuation in the tissue sample, and 4) nonlinear attenuation in the tissue sample. Furthermore, the AR model allows estimating the exponent of nonlinear attenuation. The results of this study suggest that the AR model approach can improve current QAM by providing more reliable, quantitative, tissue-property estimates and also provides additional values of parameters related to nonlinear attenuation.

Early assessment of burn severity in human tissue ex vivo with multi-wavelength spatial frequency domain imaging.

Early knowledge about burn severity and depth can lead to improved outcome for patients. In this study, we investigated the change in optical properties in ex vivo human skin following thermal burn injuries. Human skin removed during body contouring procedures was subjected to thermal burn injury for either 10 or 60 s. Multi-wavelength spatial frequency domain imaging (SFDI) measurements were performed on each sample and the optical properties (absorption and scattering parameters) were obtained at each wavelength. Multi-wavelength fitting was used to quantify absorption and scattering parameters, and these parameters were compared to histologic assessments of burn depth related to burn severity. Our results indicated substantial changes in optical scattering parameters and these changes correlated well with the burn severity and depth, and fit closely with previously reported studies using porcine in vivo models. This study provides the characterization of thermal burn injury on human skin ex vivo by using the optical method of SFDI with high sensitivity and specificity. This preclinical human model system without live animals could have uses in testing the imaging parameters of other skin injuries, including from caustic agents.

Improved High-Frequency Ultrasound Corneal Biometric Accuracy by Micrometer-Resolution Acoustic-Property Maps of the Cornea.

PURPOSE: Mapping of epithelial thickness (ET) is useful for detection of keratoconus, a disease characterized by corneal thinning and bulging in which epithelial thinning occurs over the apex. In prior clinical studies, optical coherence tomography (OCT) measurements of ET were systematically thinner than those obtained by 40-MHz high-frequency ultrasound (HFU) where a constant speed of sound (c) of 1636 m/s was used for all corneal layers. The purpose of this work was to study the acoustic properties, that is, c, acoustic impedance (Z), and attenuation (α) of the corneal epithelium and stroma independently using a scanning acoustic microscope (SAM) to investigate the discrepancy between OCT and HFU estimates of ET. METHODS: Twelve unfixed pig corneas were snap-frozen and 6-µm sections were scanned using a custom-built SAM with an F-1.08, 500-MHz transducer and a 264-MHz bandwidth. Two-dimensional maps of c, Z, and α with a spatial resolution of 4 µm were derived. RESULTS: SAM showed that the value of c in the epithelium (i.e., 1548 ± 18 m/s) is substantially lower than the value of c in the stroma (i.e., 1686 ± 33 m/s). CONCLUSION: SAM results demonstrated that the assumption of a constant value of c for all corneal layers is incorrect and explains the prior discrepancy between OCT and HFU ET determinations. TRANSLATIONAL RELEVANCE: The findings of this study have important implications for HFU-based ET measurements and will improve future keratoconus diagnosis by providing more-accurate ET estimates.

High-Frequency Quantitative Ultrasound for Imaging Prostate Cancer Using a Novel Micro-Ultrasound Scanner.

Currently, biopsies guided by transrectal ultrasound (TRUS) are the only method for definitive diagnosis of prostate cancer. Studies by our group suggest that quantitative ultrasound (QUS) could provide a more sensitive means of targeting biopsies and directing focal treatments to cancer-suspicious regions in the prostate. Previous studies have utilized ultrasound signals at typical clinical frequencies, i.e., in the 6-MHz range. In the present study, a 29-MHz, TRUS, micro-ultrasound system and transducer (ExactVu micro-ultrasound, Exact Imaging, Markham, Canada) was used to acquire radio frequency data from 163 patients immediately before 12-core biopsy procedures, comprising 1956 cores. These retrospective data are a subset of data acquired in an ongoing, multisite, 2000-patient, randomized, clinical trial (clinicaltrials.gov NCT02079025). Spectrum-based QUS estimates of effective scatter diameter (ESD), effective acoustic concentration (EAC), midband (M), intercept (I) and slope (S) as well as envelope statistics employing a Nakagami distribution were used to train linear discriminant classifiers (LDCs) and support vector machines (SVMs). Classifier performance was assessed using area-under-the-curve (AUC) values obtained from receiver operating characteristic (ROC) analyses with 10-fold cross validation. A combination of ESD and EAC parameters resulted in an AUC value of 0.77 using a LDC. When Nakagami-µ or prostate-specific antigen (PSA) values were added as features, the AUC value increased to 0.79. SVM produced an AUC value of 0.77, using a combination of envelope and spectral QUS estimates. The best classification produced an AUC value of 0.81 using an LDC when combining envelope statistics, PSA, ESD and EAC. In a previous study, B-mode-based scoring and evaluation using the PRI-MUS protocol produced a maximal AUC value of 0.74 for higher Gleason-score values (GS >7) when read by an expert. Our initial results with AUC values of 0.81 are very encouraging for developing a new, predominantly user-independent, prostate-cancer, risk-assessing tool.

Resting-state functional connectivity measured by diffuse correlation spectroscopy.

Near-infrared diffuse correlation spectroscopy (DCS) is used to record spontaneous cerebral blood flow fluctuations in the frontal cortex. Nine adult subjects participated in the experiments, in which 8-minute spontaneous fluctuations were simultaneously recorded from the left and right dorsolateral and inferior frontal regions. Resting-state functional connectivity (RSFC) was measured by the temporal correlation of the low frequency fluctuations. Our data shows the RSFC within the dorsolateral region is significantly stronger than that between the inferior and dorsolateral regions, in line with previous observations with functional near-infrared spectroscopy. This indicates that DCS is capable of investigating brain functional connectivity in terms of cerebral blood flow.

A dual-channel endoscope for quantitative imaging, monitoring, and triggering of doxorubicin release from liposomes in living mice.

Doxorubicin (Dox) is approved for use in liposomal form for the treatment of ovarian cancer. We previously developed a long-circulating Dox formulation in liposomes containing small amounts of porphyrin-phospholipid, which enables on-demand drug release with near-infrared irradiation. In this study, we present and evaluate a dual-modal, dual-channel light endoscope that allows quantitative reflectance and fluorescence imaging for monitoring of local Dox concentrations in target areas. The endoscope consists of two flexible imaging fibers; one to transmit diagnostic and therapeutic light to the target, and the other to detect fluorescent and reflected light. Thus, the endoscope serves for imaging, for light delivery to trigger drug release, and for monitoring drug concentration kinetics during drug release. We characterized the performance of this endoscope in tissue phantoms and in an in vivo model of ovarian cancer. This study demonstrates the feasibility of non-invasive, quantitative mapping of Dox distribution in vivo via endoscopic imaging.

Noninvasive mesoscopic imaging of actinic skin damage using spatial frequency domain imaging.

For prevention and accurate intervention planning, it is crucial to predict if lesions will progress towards cancer. In this study, we investigated the change in optical properties and vascular parameters to characterize skin tissue from mild photodamage to actinic keratosis (AK). Multi-wavelength spatial frequency domain imaging (SFDI) measurements were performed on three patients with clinically normal skin, as well as pre-cancerous actinic keratosis lesions. Our results indicate that there exist significant differences in both optical and vascular parameters between these patients, and that these parameters can be early biomarkers of neoplasia. Ultimately, clinicians can use this noninvasive approach for frequent monitoring of high-risk population.

Material Properties of Human Ocular Tissue at 7-µm Resolution.

Quantitative assessment of the material properties of ocular tissues can provide valuable information for investigating several ophthalmic diseases. Quantitative acoustic microscopy (QAM) offers a means of obtaining such information, but few QAM investigations have been conducted on human ocular tissue. We imaged the optic nerve (ON) and iridocorneal angle in 12-µm deparaffinized sections of the human eye using a custom-built acoustic microscope with a 250-MHz transducer (7-µm lateral resolution). The two-dimensional QAM maps of ultrasound attenuation (α), speed of sound ( c), acoustic impedance ( Z), bulk modulus ( K), and mass density (ρ) were generated. Scanned samples were then stained and imaged by light microscopy for comparison with QAM maps. The spatial resolution and contrast of scanning acoustic microscopy (SAM) maps were sufficient to resolve anatomic layers of the retina (Re); anatomic features in SAM maps corresponded to those seen by light microscopy. Significant variations of the acoustic parameters were found. For example, the sclera was 220 MPa stiffer than Re, choroid, and ON tissue. To the authors' knowledge, this is the first systematic study to assess c, Z, K, ρ, and α of human ocular tissue at the high ultrasound frequencies used in this study.

Optical imaging of tissue obtained by transbronchial biopsies of peripheral lung lesions.

BACKGROUND: Bronchoscopic procedures have been increasingly used for the diagnosis of peripheral lung cancers, but the yield remains moderately low. The aim of this study is to assess the feasibility and ability of a custom-built bimodal optical spectroscopy system to enhance the on-site discrimination between malignant and benign specimens obtained from the transbronchial lung biopsies (TBLB) of peripheral lung lesions. METHODS: We conducted a prospective and single-center pilot study to examine the TBLB specimens obtained from peripheral lung lesions. Diffuse reflectance spectroscopy (DRS) and diffuse fluorescence spectroscopy (DFS) parameters were used to analyze the optical characteristics of these specimens. RESULTS: One hundred and sixteen biopsy specimens from 15 patients were analyzed using optical imaging. All specimens had a confirmed pathologic diagnosis. Notably, 22 of the 116 specimens were malignant, and 10 of the 94 non-malignant specimens were necrotic biopsies. Individual parameters showed significant difference between the three groups (malignant, non-malignant and necrosis). Multivariate analysis of the blood, scattering and fluorescence parameters demonstrated a sensitivity of 77.3% and specificity of 73.1% in differentiating between malignant and benign specimens and a sensitivity of 90.9% and specificity of 100% in differentiating malignant from necrotic specimens. CONCLUSIONS: We conclude that optical spectroscopy is a feasible modality for on-site discrimination between malignant and benign as well as malignant and necrotic TBLB specimens of peripheral lung lesions.

Regular chondrocyte spacing is a potential cause for coherent ultrasound backscatter in human articular cartilage.

The potential of quantitative ultrasound (QUS) to assess the regular cellular spacing in the superficial cartilage zones was investigated experimentally and numerically. Nine osteochondral samples, extracted from two human cadaver knee joints, were measured using a 50-MHz ultrasound scanning device and evaluated using Mankin score. Simulated backscattered power spectra from models with an idealized cell alignment exhibited a pronounced frequency peak. From the peak, cell spacing in the range between 15 and 40 µm between cell layers was detected with an average error of 0.2 µm. The mean QUS-based cell spacing was 28.3 ± 5.3 µm. Strong correlation (R2 = 0.59, p ≤ 0.001) between spacing estimates from light microscopy (LM) and QUS was found for samples with Mankin score ≤3. For higher scores, QUS-based spacing was significantly higher (p ≤ 0.05) compared to LM-based spacing. QUS-based spacing estimates together with other QUS parameters may serve as future biomarkers for detecting early signs of osteoarthrosis.

Structural and Epimeric Isomers of HPPH [3-Devinyl 3-{1-(1-hexyloxy) ethyl}pyropheophorbide-a]: Effects on Uptake and Photodynamic Therapy of Cancer.

The tetrapyrrole structure of porphyrins used as photosentizing agents is thought to determine uptake and retention by malignant epithelial cancer cells. To assess the contribution of the oxidized state of individual rings to these cellular processes, bacteriochlorophyll a was converted into the ring "D" reduced 3-devinyl-3-[1-(1-hexyloxy)ethyl]pyropheophorbide-a (HPPH) and the corresponding ring "B" reduced isomer (iso-HPPH). The carboxylic acid analogs of both ring "B" and ring "D" reduced isomers showed several-fold higher accumulation into the mitochondria and endoplasmic reticulum by primary culture of human lung and head and neck cancer cells than the corresponding methyl ester analogs that localize primarily to granular vesicles and to a lesser extent to mitochondria. However, long-term cellular retention of these compounds exhibited an inverse relationship with tumor cells generally retaining better the methyl-ester derivatives. In vivo distribution and tumor uptake was evaluated in the isogenic model of BALB/c mice bearing Colon26 tumors using the respective 14C-labeled analogs. Both carboxylic acid derivatives demonstrated similar intracellular localization and long-term tumor cure with no significant skin phototoxicity. PDT-mediated tumor action involved vascular damage, which was confirmed by a reduction in blood flow and immunohistochemical assessment of damage to the vascular endothelium. The HPPH stereoisomers (epimers) showed identical uptake (in vitro & in vivo), intracellular retention and photoreaction.

A Novel Quantitative 500-MHz Acoustic Microscopy System for Ophthalmologic Tissues.

OBJECTIVE: This paper describes development of a novel 500-MHz scanning acoustic microscope (SAM) for assessing the mechanical properties of ocular tissues at fine resolution. The mechanical properties of some ocular tissues, such as lamina cribrosa (LC) in the optic nerve head, are believed to play a pivotal role in eye pathogenesis. METHODS: A novel etching technology was used to fabricate silicon-based lens for a 500-MHz transducer. The transducer was tested in a custom-designed scanning system on human eyes. Two-dimensional (2-D) maps of bulk modulus (K) and mass density (ρ) were derived using improved versions of current state-of-the-art signal processing approaches. RESULTS: The transducer employed a lens radius of 125 µm and had a center frequency of 479 MHz with a -6-dB bandwidth of 264 MHz and a lateral resolution of 4 µm. The LC, Bruch's membrane (BM) at the interface of the retina and choroid, and Bowman's layer (BL) at the interface of the corneal epithelium and stroma, were successfully imaged and resolved. Analysis of the 2-D parameter maps revealed average values of LC, BM, and BL with KLC = 2.81 ±0.17; GPa, KBM = 2.89 ±0.18; GPa, KBL = 2.6 ±0.09 ; GPa, ρ LC = 0.96 ±0.03 g/cm3; ρ BM = 0.97 ±0.04 g/cm3; ρ BL = 0.98 ±0.04 g/cm3. SIGNIFICANCE: This novel SAM was shown to be capable of measuring mechanical properties of soft biological tissues at microscopic resolution; it is currently the only system that allows simultaneous measurement of K, ρ, and attenuation in large lateral scales (field area >9 mm2) and at fine resolutions.

Quantitative imaging of light-triggered doxorubicin release.

The efficacy of chemotherapy is related, in large part, to the concentration of drug that reaches tumor sites. Doxorubicin (DOX) is a common anti-cancer drug that is also approved for use in liposomal form for the treatment of ovarian cancer. We recently developed a porphyrin-phospholipid (PoP)-liposome system that enables on demand release of DOX from liposomes using near infrared irradiation to improve DOX bioavailability. Owing to its intrinsic fluorescence, it is possible, and desirable, to quantify DOX concentration and distribution, preferably noninvasively. Here we quantified DOX distribution following light-triggered drug release in phantoms and an animal carcass using spatial frequency domain imaging. This study demonstrates the feasibility of non-invasive quantitative mapping of DOX distributions in target areas.

Characterization of nonmelanoma skin cancer for light therapy using spatial frequency domain imaging.

The dosimetry of light-based therapies critically depends on both optical and vascular parameters. We utilized spatial frequency domain imaging to quantify optical and vascular parameters, as well as estimated light penetration depth from 17 nonmelanoma skin cancer patients. Our data indicates that there exist substantial spatial variations in these parameters. Characterization of these parameters may inform understanding and optimization of the clinical response of light-based therapies.