David M. McClatchy

Sub-diffusive scattering parameter maps recovered using wide-field high-frequency structured light imaging

This study investigates the hypothesis that structured light reflectance imaging with high spatial frequency patterns [Formula: see text] can be used to quantitatively map the anisotropic scattering phase function distribution [Formula: see text] in turbid media. Monte Carlo simulations were used in part to establish a semi-empirical model of demodulated reflectance ([Formula: see text]) in terms of dimensionless scattering [Formula: see text] and [Formula: see text], a metric of the first two moments of the [Formula: see text] distribution. Experiments completed in tissue-simulating phantoms showed that simultaneous analysis of [Formula: see text] spectra sampled at multiple [Formula: see text] in the frequency range [0.05-0.5] [Formula: see text] allowed accurate estimation of both [Formula: see text] in the relevant tissue range [0.4-1.8] [Formula: see text], and [Formula: see text] in the range [1.4-1.75]. Pilot measurements of a healthy volunteer exhibited [Formula: see text]-based contrast between scar tissue and surrounding normal skin, which was not as apparent in wide field diffuse imaging. These results represent the first wide-field maps to quantify sub-diffuse scattering parameters, which are sensitive to sub-microscopic tissue structures and composition, and therefore, offer potential for fast diagnostic imaging of ultrastructure on a size scale that is relevant to surgical applications.

Projection imaging of photon beams by the Čerenkov effect

PURPOSE A novel technique for beam profiling of megavoltage photon beams was investigated for the first time by capturing images of the induced Čerenkov emission in water, as a potential surrogate for the imparted dose in irradiated media. METHODS A high-sensitivity, intensified CCD camera (ICCD) was configured to acquire 2D projection images of Čerenkov emission from a 4 × 4 cm(2) 6 MV linear accelerator (LINAC) x-ray photon beam operating at a dose rate of 400 MU∕min incident on a water tank with transparent walls. The ICCD acquisition was gated to the LINAC sync pulse to reduce background light artifacts, and the measurement quality was investigated by evaluating the signal to noise ratio and measurement repeatability as a function of delivered dose. Monte Carlo simulations were used to derive a calibration factor for differences between the optical images and deposited dose arising from the anisotropic angular dependence of Čerenkov emission. Finally, Čerenkov-based beam profiles were compared to a percent depth dose (PDD) and lateral dose profile at a depth of d(max) from a reference dose distribution generated from the clinical Varian ECLIPSE treatment planning system (TPS). RESULTS The signal to noise ratio was found to be 20 at a delivered dose of 66.6 cGy, and proportional to the square root of the delivered dose as expected from Poisson photon counting statistics. A 2.1% mean standard deviation and 5.6% maximum variation in successive measurements were observed, and the Monte Carlo derived calibration factor resulted in Čerenkov emission images which were directly correlated to deposited dose, with some spatial issues. The dose difference between the TPS and PDD predicted by Čerenkov measurements was within 20% in the buildup region with a distance to agreement (DTA) of 1.5-2 mm and ±3% at depths beyond d(max). In the lateral profile, the dose difference at the beam penumbra was within ±13% with a DTA of 0-2 mm, ±5% in the central beam region, and 2%-3% in the beam umbra. CONCLUSIONS The results from this initial study demonstrate the first documented use of Čerenkov emission imaging to profile x-ray photon LINAC beams in water. The proposed modality has several potential advantages over alternative methods, and upon future refinement may prove to be a robust and novel dosimetry method.

Pulsed-light imaging for fluorescence guided surgery under normal room lighting.

Fluorescence guided surgery (FGS) is an emerging technology that has demonstrated improved surgical outcomes. However, dim lighting conditions required by current FGS systems are disruptive to standard surgical workflow. We present a novel FGS system capable of imaging fluorescence under normal room light by using pulsed excitation and gated acquisition. Images from tissue-simulating phantoms confirm visual detection down to 0.25 μM of protoporphyrin IX under 125 μW/cm2 of ambient light, more than an order of magnitude lower than that measured with the Zeiss Pentero in the dark. Resection of orthotopic brain tumors in mice also suggests that the pulsed-light system provides superior sensitivity in vivo.

Structured light scatteroscopy.

Abstract. A new imaging approach, structured light scatteroscopy (SLS), is demonstrated, which offers rapid wide-field imaging of microscopic morphological variations in bulk tissue surfaces. Elastic scattering of light offers exquisite sensitivity to ultrastructural changes at multiple size scales ranging from nanometers to millimeters, but in bulk tissues the confounding effects of molecular absorption and strong multiple scattering of light often lead to a dramatic reduction in scatter contrast and specificity. It is demonstrated that the SLS using structured high spatial frequency illumination and detection to probe the tissue achieves direct, absorption-independent, high-resolution maps of the scattering response. The scattering response is observed to be dependent on both the wavelength and spatial frequency of choice, indicating a potential for multiscale probing of ultrastructural changes in superficial tissue layers. This methodology can be easily applied in most wide-field imaging systems.

Wide-field quantitative imaging of tissue microstructure using sub-diffuse spatial frequency domain imaging

Localized measurements of scattering in biological tissue provide sensitivity to microstructural morphology but have limited utility to wide-field applications, such as surgical guidance. This study introduces sub-diffusive spatial frequency domain imaging (sd-SFDI), which uses high spatial frequency illumination to achieve wide-field sampling of localized reflectances. Model-based inversion recovers macroscopic variations in the reduced scattering coefficient [Formula: see text] and the phase function backscatter parameter (γ). Measurements in optical phantoms show quantitative imaging of user-tuned phase-function-based contrast with accurate decoupling of parameters that define both the density and the size-scale distribution of scatterers. Measurements of fresh ex vivo breast tissue samples revealed, for the first time, unique clustering of sub-diffusive scattering properties for different tissue types. The results support that sd-SFDI provides maps of microscopic structural biomarkers that cannot be obtained with diffuse wide-field imaging and characterizes spatial variations not resolved by point-based optical sampling.

Molecular dyes used for surgical specimen margin orientation allow for intraoperative optical assessment during breast conserving surgery

A variety of optical techniques utilizing near-infrared (NIR) light are being proposed for intraoperative breast tumor margin assessment. However, immediately following a lumpectomy excision, the margins are inked, which preserves the orientation of the specimen but prevents optical interrogation of the tissue margins. Here, a workflow is proposed that allows for both NIR optical assessment following full specimen marking using molecular dyes which have negligible absorption and scattering in the NIR. The effect of standard surgical inks in contrast to molecular dyes for an NIR signal is shown. Further, the proposed workflow is demonstrated with full specimen intraoperative imaging on all margins directly after the lumpectomy has been excised and completely marked. This work is an important step in the path to clinical feasibility of intraoperative breast tumor margin assessment using NIR optical methods without having to compromise on the current clinical practice of inking resected specimens for margin orientation.

Calibration and analysis of a multimodal micro-CT and structured light imaging system for the evaluation of excised breast tissue

A multimodal micro-computed tomography (CT) and multi-spectral structured light imaging (SLI) system is introduced and systematically analyzed to test its feasibility to aid in margin delineation during breast conserving surgery (BCS). Phantom analysis of the micro-CT yielded a signal-to-noise ratio of 34, a contrast of 1.64, and a minimum detectable resolution of 240 μm for a 1.2 min scan. The SLI system, spanning wavelengths 490 nm to 800 nm and spatial frequencies up to 1.37 [Formula: see text], was evaluated with aqueous tissue simulating phantoms having variations in particle size distribution, scatter density, and blood volume fraction. The reduced scattering coefficient, [Formula: see text] and phase function parameter, γ, were accurately recovered over all wavelengths independent of blood volume fractions from 0% to 4%, assuming a flat sample geometry perpendicular to the imaging plane. The resolution of the optical system was tested with a step phantom, from which the modulation transfer function was calculated yielding a maximum resolution of 3.78 cycles per mm. The three dimensional spatial co-registration between the CT and optical imaging space was tested and shown to be accurate within 0.7 mm. A freshly resected breast specimen, with lobular carcinoma, fibrocystic disease, and adipose, was imaged with the system. The micro-CT provided visualization of the tumor mass and its spiculations, and SLI yielded superficial quantification of light scattering parameters for the malignant and benign tissue types. These results appear to be the first demonstration of SLI combined with standard medical tomography for imaging excised tumor specimens. While further investigations are needed to determine and test the spectral, spatial, and CT features required to classify tissue, this study demonstrates the ability of multimodal CT/SLI to quantify, visualize, and spatially navigate breast tumor specimens, which could potentially aid in the assessment of tumor margin status during BCS.

High spatial frequency structured light imaging for intraoperative breast tumor margin assessment

A new superficial wide-field imaging technique is presented, which utilizes high spatial frequency structured illumination to constrain the light sampling volume to a sub-diffuse regime. In this transport regime, the effects of absorption are drastically reduced and the sensitivity to local scattering from ultrastructrual alterations is increased. Absorption independence is validated with multiple experiments including a bovine blood-Intralipid solution matrix and avian tissue with superficial bovine blood. The resulting structured light demodulated images show a complete insensitivity to the blood over Hb concentrations of 0 – 240 μM. Increased sensitivity to ultrastructual changes is demonstrated by imaging avian tissue with controlled morphological alterations including formalin-induced crosslinking. This imaging technique is currently being translated towards intraoperative assessment of breast tumor margins because of its ability to capture an entire lumpectomy margin in a single field of view, insensitivity to confounding surface blood present on lumpectomies, and its inherent scatter signal without the need of any model inversion. Further, a new lumpectomy marking system is introduced that allows for both the surgeon to mark the lumpectomy during excision and optical assessment of the specimen after the margins have been marked. Structured light images acquired intraoperatively of all margins of lumpectomy specimens are presented to show feasibility of clinical translation. Immediate future work will focus on developing a multi-spectral system.

Wide-field quantitative imaging of intrinsic scatter bio-markers using sub-diffusive structured light

Sub-diffusive structured light imaging can quantitate the density of scatterers versus their size scale distribution in a wide-field geometry. Phantoms with unique fractal distributions and n=22 fresh human breast specimens are imaged and analyzed.

High spatial frequency structured light imaging negates absorption effects in fluorescence imaging

Fluorescence guided surgery is hampered by absorption and scatter, which confound interpretation of fluorescence. High-frequency fluorescence structured light imaging enables real-time model-independent absorption and scatter correction.