Intraoperative optical coherence tomography for assessing human lymph nodes for metastatic cancer.

BACKGROUND: Evaluation of lymph node (LN) status is an important factor for detecting metastasis and thereby staging breast cancer. Currently utilized clinical techniques involve the surgical disruption and resection of lymphatic structure, whether nodes or axillary contents, for histological examination. While reasonably effective at detection of macrometastasis, the majority of the resected lymph nodes are histologically negative. Improvements need to be made to better detect micrometastasis, minimize or eliminate lymphatic disruption complications, and provide immediate and accurate intraoperative feedback for in vivo cancer staging to better guide surgery. METHODS: We evaluated the use of optical coherence tomography (OCT), a high-resolution, real-time, label-free imaging modality for the intraoperative assessment of human LNs for metastatic disease in patients with breast cancer. We assessed the sensitivity and specificity of double-blinded trained readers who analyzed intraoperative OCT LN images for presence of metastatic disease, using co-registered post-operative histopathology as the gold standard. RESULTS: Our results suggest that intraoperative OCT examination of LNs is an appropriate real-time, label-free, non-destructive alternative to frozen-section analysis, potentially offering faster interpretation and results to empower superior intraoperative decision-making. CONCLUSIONS: Intraoperative OCT has strong potential to supplement current post-operative histopathology with real-time in situ assessment of LNs to preserve both non-cancerous nodes and their lymphatic vessels, and thus reduce the associated risks and complications from surgical disruption of lymphoid structures following biopsy.

Stability in computed optical interferometric tomography (part I): stability requirements.

As imaging systems become more advanced and acquire data at faster rates, increasingly dynamic samples can be imaged without concern of motion artifacts. For optical interferometric techniques such as optical coherence tomography, it often follows that initially, only amplitude-based data are utilized due to unstable or unreliable phase measurements. As systems progress, stable phase maps can also be acquired, enabling more advanced, phase-dependent post-processing techniques. Here we report an investigation of the stability requirements for a class of phase-dependent post-processing techniques - numerical defocus and aberration correction with further extensions to techniques such as Doppler, phase-variance, and optical coherence elastography. Mathematical analyses and numerical simulations over a variety of instabilities are supported by experimental investigations.

Stability in computed optical interferometric tomography (Part II): in vivo stability assessment.

Stability is of utmost importance to a wide range of phase-sensitive processing techniques. In Doppler optical coherence tomography and optical coherence elastography, in addition to defocus and aberration correction techniques such as interferometric synthetic aperture microscopy and computational/digital adaptive optics, a precise understanding of the system and sample stability helps to guide the system design and choice of imaging parameters. This article focuses on methods to accurately and quantitatively measure the stability of an imaging configuration in vivo. These methods are capable of partially decoupling axial from transverse motion and are compared against the stability requirements for computed optical interferometric tomography laid out in the first part of this article.

Quantification of Retinal Microvascular Density in Optical Coherence Tomographic Angiography Images in Diabetic Retinopathy.

Importance: Quantitative measurements based on optical coherence tomographic angiography (OCTA) may have value in managing diabetic retinopathy (DR), but there is limited information on the ability of OCTA to distinguish eyes with DR. Objective: To evaluate the ability of measurements of retinal microvasculature using OCTA to distinguish healthy eyes from eyes with DR. Design, Setting, and Participants: In this prospective cross-sectional study, OCTA was used to examine the eyes of participants with type 2 diabetes with or without DR and the eyes of participants without diabetes from September 17, 2015, to April 6, 2016. Density maps based on superficial retinal layer (SRL) and deeper retinal layer (DRL) images were generated after a method to remove decorrelation tails was applied to the DRL images. Exposures: Both eyes of each participant were examined by means of a 3-mm OCTA scan and 7-field fundus photography using the Diabetic Retinopathy Severity Scale. Main Outcomes and Measures: Two measures were examined: perfusion density, based on the area of vessels, and vessel density, based on a map with vessels of 1-pixel width. The size of the foveal avascular zone was also calculated automatically, and so was the area under the receiver operating characteristic curve. Results: A total of 50 eyes from 26 participants with diabetes (10 women and 16 men; mean [SD] age, 64.9 [7.5] years) and 50 healthy eyes from 25 participants without diabetes (14 women and 11 men; mean [SD] age, 64.0 [7.1] years) were imaged. All participants were white. Vessel density measured in the SRL had the highest area under the receiver operating characteristic curve (0.893 [95% CI, 0.827-0.959]), compared with perfusion density in the SRL (0.794 [95% CI, 0.707-0.881]), foveal avascular zone area (0.472 [95% CI, 0.356-0.588]), and vessel density in the DRL (0.703 [95% CI, 0.601-0.805]). Vessel density in the SRL negatively correlated with best-corrected visual acuity (r = -0.28; P = .05) and severity of DR (r = -0.46; P = .001). Density metrics correlated with age. No correlation was detected between vascular density or foveal avascular zone metrics and hemoglobin A1C or duration of diabetes. Conclusions and Relevance: Vessel density measured by OCTA provides a quantitative metric of capillary closure that correlates with severity of DR and may allow staging, diagnosis, and monitoring that do not require subjective evaluation of fundus images.

Optical Coherence Tomography Angiography Macular and Peripapillary Vessel Perfusion Density in Healthy Subjects, Glaucoma Suspects, and Glaucoma Patients.

Purpose: To evaluate macular and peripapillary vessel perfusion density (VD) in glaucoma suspects (GS) and glaucoma patients; to correlate ganglion cell-inner plexiform layer (GCIPL) and retinal nerve fiber layer (RNFL) thicknesses with macular and peripapillary VD; and to evaluate the diagnostic accuracy of the structural and vascular parameters. Methods: A consecutive series of GS, glaucoma patients, and healthy subjects was prospectively recruited from July 1, 2016, to January 31, 2017. All subjects underwent standard automated perimetry, spectral-domain optical coherence tomography (OCT), and 6 × 6-mm optical coherence tomography angiography (OCT-A) centered on the fovea and optic nerve. Results: Forty controls, 40 GS, and 40 glaucoma patients were enrolled. Peripapillary RNFL, GCIPL, and macular RNFL thicknesses significantly decreased in the glaucoma group compared to controls and GS (P < 0.01). Peripapillary VD in average and in the superior and inferior quadrants decreased in the glaucoma group (P ≤ 0.001); conversely, macular VD was not statistically different across groups (P > 0.05). At the peripapillary area, a correlation between RNFL thickness and VD was found; conversely, no statistically significant correlation was found between GCIPL thicknesses and macular VD (all P > 0.05) in all groups. Peripapillary RNFL and GCIPL showed higher diagnostic capacity compared to peripapillary and macular VDs. Conclusions: Structural damage is evident both in the peripapillary and in macular areas. Vascular damage seems to be less prominent, as it was seen only for the glaucoma group and at the radial peripapillary plexus. Diagnostic abilities are excellent for structural variables, less so but still good for peripapillary VD, and poor for macular VD.

Computational high-resolution optical imaging of the living human retina.

High-resolution in vivo imaging is of great importance for the fields of biology and medicine. The introduction of hardware-based adaptive optics (HAO) has pushed the limits of optical imaging, enabling high-resolution near diffraction-limited imaging of previously unresolvable structures1,2. In ophthalmology, when combined with optical coherence tomography, HAO has enabled a detailed three-dimensional visualization of photoreceptor distributions3,4 and individual nerve fibre bundles5 in the living human retina. However, the introduction of HAO hardware and supporting software adds considerable complexity and cost to an imaging system, limiting the number of researchers and medical professionals who could benefit from the technology. Here we demonstrate a fully automated computational approach that enables high-resolution in vivo ophthalmic imaging without the need for HAO. The results demonstrate that computational methods in coherent microscopy are applicable in highly dynamic living systems.

Polarization-sensitive interferometric synthetic aperture microscopy.

Three-dimensional optical microscopy suffers from the well-known compromise between transverse resolution and depth-of-field. This is true for both structural imaging methods and their functional extensions. Interferometric synthetic aperture microscopy (ISAM) is a solution to the 3D coherent microscopy inverse problem that provides depth-independent transverse resolution. We demonstrate the extension of ISAM to polarization sensitive imaging, termed polarization-sensitive interferometric synthetic aperture microscopy (PS-ISAM). This technique is the first functionalization of the ISAM method and provides improved depth-of-field for polarization-sensitive imaging. The basic assumptions of polarization-sensitive imaging are explored, and refocusing of birefringent structures is experimentally demonstrated. PS-ISAM enables high-resolution volumetric imaging of birefringent materials and tissue.

Real-time automated thickness measurement of the in vivo human tympanic membrane using optical coherence tomography.

BACKGROUND: Otitis media (OM), an infection in the middle ear, is extremely common in the pediatric population. Current gold-standard methods for diagnosis include otoscopy for visualizing the surface features of the tympanic membrane (TM) and making qualitative assessments to determine middle ear content. OM typically presents as an acute infection, but can progress to chronic OM, and after numerous infections and antibiotic treatments over the course of many months, this disease is often treated by surgically inserting small tubes in the TM to relieve pressure, enable drainage, and provide aeration to the middle ear. Diagnosis and monitoring of OM is critical for successful management, but remains largely qualitative. METHODS: We have developed an optical coherence tomography (OCT) system for high-resolution, depth-resolved, cross-sectional imaging of the TM and middle ear content, and for the quantitative assessment of in vivo TM thickness including the presence or absence of a middle ear biofilm. A novel algorithm was developed and demonstrated for automatic, real-time, and accurate measurement of TM thickness to aid in the diagnosis and monitoring of OM and other middle ear conditions. The segmentation algorithm applies a Hough transform to the OCT image data to determine the boundaries of the TM to calculate thickness. RESULTS: The use of OCT and this segmentation algorithm is demonstrated first on layered phantoms and then during real-time acquisition of in vivo OCT from humans. For the layered phantoms, measured thicknesses varied by approximately 5 µm over time in the presence of large axial and rotational motion. In vivo data also demonstrated differences in thicknesses both spatially on a single TM, and across normal, acute, and chronic OM cases. CONCLUSIONS: Real-time segmentation and thickness measurements of image data from both healthy subjects and those with acute and chronic OM demonstrate the use of OCT and this algorithm as a robust, quantitative, and accurate method for use during real-time in vivo human imaging.

Real-time Imaging of the Resection Bed Using a Handheld Probe to Reduce Incidence of Microscopic Positive Margins in Cancer Surgery.

Wide local excision (WLE) is a common surgical intervention for solid tumors such as those in melanoma, breast, pancreatic, and gastrointestinal cancer. However, adequate margin assessment during WLE remains a significant challenge, resulting in surgical reinterventions to achieve adequate local control. Currently, no label-free imaging method is available for surgeons to examine the resection bed in vivo for microscopic residual cancer. Optical coherence tomography (OCT) enables real-time high-resolution imaging of tissue microstructure. Previous studies have demonstrated that OCT analysis of excised tissue specimens can distinguish between normal and cancerous tissues by identifying the heterogeneous and disorganized microscopic tissue structures indicative of malignancy. In this translational study involving 35 patients, a handheld surgical OCT imaging probe was developed for in vivo use to assess margins both in the resection bed and on excised specimens for the microscopic presence of cancer. The image results from OCT showed structural differences between normal and cancerous tissue within the resection bed following WLE of the human breast. The ex vivo images were compared with standard postoperative histopathology to yield sensitivity of 91.7% [95% confidence interval (CI), 62.5%-100%] and specificity of 92.1% (95% CI, 78.4%-98%). This study demonstrates in vivo OCT imaging of the resection bed during WLE with the potential for real-time microscopic image-guided surgery.

Volumetric full-range magnetomotive optical coherence tomography.

Magnetomotive optical coherence tomography (MM-OCT) can be utilized to spatially localize the presence of magnetic particles within tissues or organs. These magnetic particle-containing regions are detected by using the capability of OCT to measure small-scale displacements induced by the activation of an external electromagnet coil typically driven by a harmonic excitation signal. The constraints imposed by the scanning schemes employed and tissue viscoelastic properties limit the speed at which conventional MM-OCT data can be acquired. Realizing that electromagnet coils can be designed to exert MM force on relatively large tissue volumes (comparable or larger than typical OCT imaging fields of view), we show that an order-of-magnitude improvement in three-dimensional (3-D) MM-OCT imaging speed can be achieved by rapid acquisition of a volumetric scan during the activation of the coil. Furthermore, we show volumetric (3-D) MM-OCT imaging over a large imaging depth range by combining this volumetric scan scheme with full-range OCT. Results with tissue equivalent phantoms and a biological tissue are shown to demonstrate this technique.

Magnetomotive optical coherence elastography using magnetic particles to induce mechanical waves.

Magnetic particles are versatile imaging agents that have found wide spread applicability in diagnostic, therapeutic, and rheology applications. In this study, we demonstrate that mechanical waves generated by a localized inclusion of magnetic nanoparticles can be used for assessment of the tissue viscoelastic properties using magnetomotive optical coherence elastography. We show these capabilities in tissue mimicking elastic and viscoelastic phantoms and in biological tissues by measuring the shear wave speed under magnetomotive excitation. Furthermore, we demonstrate the extraction of the complex shear modulus by measuring the shear wave speed at different frequencies and fitting to a Kelvin-Voigt model.

Three-dimensional motion correction using speckle and phase for in vivo computed optical interferometric tomography.

Over the years, many computed optical interferometric techniques have been developed to perform high-resolution volumetric tomography. By utilizing the phase and amplitude information provided with interferometric detection, post-acquisition corrections for defocus and optical aberrations can be performed. The introduction of the phase, though, can dramatically increase the sensitivity to motion (most prominently along the optical axis). In this paper, we present two algorithms which, together, can correct for motion in all three dimensions with enough accuracy for defocus and aberration correction in computed optical interferometric tomography. The first algorithm utilizes phase differences within the acquired data to correct for motion along the optical axis. The second algorithm utilizes the addition of a speckle tracking system using temporally- and spatially-coherent illumination to measure motion orthogonal to the optical axis. The use of coherent illumination allows for high-contrast speckle patterns even when imaging apparently uniform samples or when highly aberrated beams cannot be avoided.

Computed optical interferometric tomography for high-speed volumetric cellular imaging.

Three-dimensional high-resolution imaging methods are important for cellular-level research. Optical coherence microscopy (OCM) is a low-coherence-based interferometry technology for cellular imaging with both high axial and lateral resolution. Using a high-numerical-aperture objective, OCM normally has a shallow depth of field and requires scanning the focus through the entire region of interest to perform volumetric imaging. With a higher-numerical-aperture objective, the image quality of OCM is affected by and more sensitive to aberrations. Interferometric synthetic aperture microscopy (ISAM) and computational adaptive optics (CAO) are computed imaging techniques that overcome the depth-of-field limitation and the effect of optical aberrations in optical coherence tomography (OCT), respectively. In this work we combine OCM with ISAM and CAO to achieve high-speed volumetric cellular imaging. Experimental imaging results of ex vivo human breast tissue, ex vivo mouse brain tissue, in vitro fibroblast cells in 3D scaffolds, and in vivo human skin demonstrate the significant potential of this technique for high-speed volumetric cellular imaging.

Dynamic method of optical coherence elastography in determining viscoelasticity of polymers and tissues.

In this paper, we report on a novel quantitative elastography technique that combines optical coherence tomography (OCT) with acoustic radiation force (ARF) excitation to estimate the complex modulus. Sinusoidally modulated ARF excitations between 200 - 4000 Hz generate a surface wave at the tissue surface that can be related to bulk viscoelastic (VE) properties in a manner that is both precise and quantitative. This method is very well suited to studying media at high spatial resolution and over a very broad range of force frequencies. Mechanical characterization was calibrated using hydropolymers before studying liver samples. Fresh porcine liver samples were measured over time with and without formalin fixation. These data were used to evaluate the utility of the Kelvin-Voigt rheological model commonly used to fit dispersion data when estimating modulus values. We also investigated use of square-wave force excitation to measure the step response of tissues.

Real-time in vivo computed optical interferometric tomography.

High-resolution real-time tomography of scattering tissues is important for many areas of medicine and biology1-6. However, the compromise between transverse resolution and depth-of-field in addition to low sensitivity deep in tissue continue to impede progress towards cellular-level volumetric tomography. Computed imaging has the potential to solve these long-standing limitations. Interferometric synthetic aperture microscopy (ISAM)7-9 is a computed imaging technique enabling high-resolution volumetric tomography with spatially invariant resolution. However, its potential for clinical diagnostics remains largely untapped since full volume reconstructions required lengthy postprocessing, and the phase-stability requirements have been difficult to satisfy in vivo. Here we demonstrate how 3-D Fourier-domain resampling, in combination with high-speed optical coherence tomography (OCT), can achieve high-resolution in vivo tomography. Enhanced depth sensitivity was achieved over a depth-of-field extended in real time by more than an order of magnitude. This work lays the foundation for high-speed volumetric cellular-level tomography.

Real-time three-dimensional optical coherence tomography image-guided core-needle biopsy system.

Advances in optical imaging modalities, such as optical coherence tomography (OCT), enable us to observe tissue microstructure at high resolution and in real time. Currently, core-needle biopsies are guided by external imaging modalities such as ultrasound imaging and x-ray computed tomography (CT) for breast and lung masses, respectively. These image-guided procedures are frequently limited by spatial resolution when using ultrasound imaging, or by temporal resolution (rapid real-time feedback capabilities) when using x-ray CT. One feasible approach is to perform OCT within small gauge needles to optically image tissue microstructure. However, to date, no system or core-needle device has been developed that incorporates both three-dimensional OCT imaging and tissue biopsy within the same needle for true OCT-guided core-needle biopsy. We have developed and demonstrate an integrated core-needle biopsy system that utilizes catheter-based 3-D OCT for real-time image-guidance for target tissue localization, imaging of tissue immediately prior to physical biopsy, and subsequent OCT imaging of the biopsied specimen for immediate assessment at the point-of-care. OCT images of biopsied ex vivo tumor specimens acquired during core-needle placement are correlated with corresponding histology, and computational visualization of arbitrary planes within the 3-D OCT volumes enables feedback on specimen tissue type and biopsy quality. These results demonstrate the potential for using real-time 3-D OCT for needle biopsy guidance by imaging within the needle and tissue during biopsy procedures.

Guide-star-based computational adaptive optics for broadband interferometric tomography.

We present a method for the numerical correction of optical aberrations based on indirect sensing of the scattered wavefront from point-like scatterers ("guide stars") within a three-dimensional broadband interferometric tomogram. This method enables the correction of high-order monochromatic and chromatic aberrations utilizing guide stars that are revealed after numerical compensation of defocus and low-order aberrations of the optical system. Guide-star-based aberration correction in a silicone phantom with sparse sub-resolution-sized scatterers demonstrates improvement of resolution and signal-to-noise ratio over a large isotome. Results in highly scattering muscle tissue showed improved resolution of fine structure over an extended volume. Guide-star-based computational adaptive optics expands upon the use of image metrics for numerically optimizing the aberration correction in broadband interferometric tomography, and is analogous to phase-conjugation and time-reversal methods for focusing in turbid media.