Showing papers by "Stephen A. Boppart published in 2015"

Computational high-resolution optical imaging of the living human retina

Abstract: A computational optical imaging scheme is able to image the human retina without the need for any adaptive optics hardware.

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

Abstract: 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.

Intraoperative Assessment of Final Margins with a Handheld Optical Imaging Probe During Breast-Conserving Surgery May Reduce the Reoperation Rate: Results of a Multicenter Study

Abstract: Background. A multicenter, prospective, blinded study was performed to test the feasibility of using a handheld optical imaging probe for the intraoperative assessment of final surgical margins during breast-conserving surgery (BCS) and to determine the potential impact on patient outcomes. Methods. Forty-six patients with early-stage breast cancer (one with bilateral disease) undergoing BCS at two study sites, the Johns Hopkins Hospital and Anne Arundel Medical Center, were enrolled in this study. During BCS, cavity-shaved margins were obtained and the final margins were examined ex vivo in the operating room with a probe incorporating optical coherence tomography (OCT) hardware and interferometric synthetic aperture microscopy (ISAM) image processing. Images were interpreted after BCS by three physicians blinded to final pathology-reported margin status. Individual and combined interpretations were assessed. Results were compared to conventional postoperative histopathology. Results. A total of 2,191 images were collected and interpreted from 229 shave margin specimens. Of the eight patients (17 %) with positive margins (0 mm), which included invasive and in situ diseases, the device identified all positive margins in five (63 %) of them; reoperation could potentially have been avoided in these patients. Among patients with pathologically negative margins ([0 mm), an estimated mean additional tissue volume of 10.7 ml (approximately 1 % of overall breast volume) would have been unnecessarily removed due to false positives.

Role of biofilm roughness and hydrodynamic conditions in Legionella pneumophila adhesion to and detachment from simulated drinking water biofilms.

Abstract: Biofilms in drinking water distribution systems (DWDS) could exacerbate the persistence and associated risks of pathogenic Legionella pneumophila (L. pneumophila), thus raising human health concerns. However, mechanisms controlling adhesion and subsequent detachment of L. pneumophila associated with biofilms remain unclear. We determined the connection between L. pneumophila adhesion and subsequent detachment with biofilm physical structure characterization using optical coherence tomography (OCT) imaging technique. Analysis of the OCT images of multispecies biofilms grown under low nutrient condition up to 34 weeks revealed the lack of biofilm deformation even when these biofilms were exposed to flow velocity of 0.7 m/s, typical flow for DWDS. L. pneumophila adhesion on these biofilm under low flow velocity (0.007 m/s) positively correlated with biofilm roughness due to enlarged biofilm surface area and local flow conditions created by roughness asperities. The preadhered L. pneumophila on selected rough...

Noninvasive depth‐resolved optical measurements of the tympanic membrane and middle ear for differentiating otitis media

Abstract: Objective/Hypothesis In this study, optical coherence tomography (OCT) is used to noninvasively and quantitatively determine tympanic membrane (TM) thickness and the presence and thickness of any middle-ear biofilm located behind the TM. These new metrics offer the potential to differentiate normal, acute, and chronic otitis media (OM) infections in pediatric subjects. Study Design Case series with comparison group. Methods The TM thickness of 34 pediatric subjects was acquired using a custom-built, handheld OCT system following a traditional otoscopic ear exam. Results Overall thickness (TM and any associated biofilm) was shown to be statistically different for normal, acute, and chronic infection groups (normal-acute and normal-chronic: P value < 0.001; acute-chronic: P value = 0.0016). Almost all observed scans from the chronic group had an accompanying biofilm structure. When the thickness of the TM and biofilm were considered separately in chronic OM, the chronic TM thickness correlated with the normal group (P value = 0.68) yet was still distinct from the acute OM group (P value < 0.001), indicating that the TM in chronic OM returns to relatively normal thickness levels. Conclusion Identifying these physical changes in vivo provides new metrics for noninvasively and quantitatively differentiating normal, acute, and chronic OM. This new diagnostic information has the potential to assist physicians to more effectively and efficiently screen, manage, and refer patients based on quantitative data. Level of Evidence 4. Laryngoscope, 125:E276–E282, 2015

Suppressing Short-term Polarization Noise and Related Spectral Decoherence in All-normal Dispersion Fiber Supercontinuum Generation.

Abstract: The supercontinuum generated exclusively in the normal dispersion regime of a nonlinear fiber is widely believed to possess low optical noise and high spectral coherence. The recent development of flattened all-normal dispersion fibers has been motivated by this belief to construct a general-purpose broadband coherent optical source. Somewhat surprisingly, we identify a large short-term polarization noise in this type of supercontinuum generation that has been masked by the total-intensity measurement in the past, but can be easily detected by filtering the supercontinuum with a linear polarizer. Fortunately, this hidden intrinsic noise and the accompanied spectral decoherence can be effectively suppressed by using a polarization-maintaining all-normal dispersion fiber. A polarization-maintaining coherent supercontinuum laser is thus built with a broad bandwidth (780-1300 nm) and high spectral power (~1 mW/nm).

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

Abstract: 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.

Optical Coherence Elastography

Abstract: OCE provides 3-D maps of tissue stiffness on the micrometer to millimeter scale, bridging a crucial gap in probing the mechanical properties of diseased tissues.

Effect of recombinant interleukin-12 on murine skin regeneration and cell dynamics using in vivo multimodal microscopy.

Abstract: Interleukin-12 (IL-12) is a pro-inflammatory cytokine known for its role in immunity, and previous studies have shown that IL-12 provides mitigation of radiation injury. In this study, we utilize a multimodal microscopy system equipped with second harmonic generation (SHG) and fluorescence lifetime imaging microscopy (FLIM) to examine the effect of IL-12 on collagen structure and cellular metabolic activity in vivo during skin wound healing. This preliminary study illustrates the highly dynamic and heterogeneous in vivo microenvironment of the wounded skin. In addition, results suggest that IL-12 triggers a significantly more rapid and greater cellular metabolic response in the wounded animals. These results can elucidate insights into the response mechanism of IL-12 in both wound healing and acute radiation syndrome.

Mechanical contrast in spectroscopic magnetomotive optical coherence elastography.

Abstract: The viscoelastic properties of tissues are altered during pathogenesis of numerous diseases and can therefore be a useful indicator of disease status and progression. Several elastography studies have utilized the mechanical frequency response and the resonance frequencies of tissue samples to characterize their mechanical properties. However, using the resonance frequency as a source of mechanical contrast in heterogeneous samples is complicated because it not only depends on the viscoelastic properties but also on the geometry and boundary conditions. In an elastography technique called magnetomotive optical coherence elastography (MM-OCE), the controlled movement of magnetic nanoparticles (MNPs) within the sample is used to obtain the mechanical properties. Previous demonstrations of MM-OCE have typically used point measurements in elastically homogeneous samples assuming a uniform concentration of MNPs. In this study, we evaluate the feasibility of generating MM-OCE elastograms in heterogeneous samples based on a spectroscopic approach which involves measuring the magnetomotive response at different excitation frequencies. Biological tissues and tissue-mimicking phantoms with two elastically distinct regions placed in side-by-side and bilayer configurations were used for the experiments, and finite element method simulations were used to validate the experimental results.

Enhancement and wavelength-shifted emission of Cerenkov luminescence using multifunctional microspheres.

Abstract: Cerenkov luminescence (CL) imaging is a new molecular imaging modality that utilizes the photons emitted during radioactive decay when charged particles travel faster than the phase velocity of light in a dielectric medium. Here we present a novel agent to convert and increase CL emission at longer wavelengths using multimodal protein microspheres (MSs). The 64Cu-labeled protein microspheres contain quantum dots (QDs) encapsulated within a high-refractive-index-oil core. Dark box imaging of the MSs was conducted to demonstrate the improvement in CL emission at longer wavelengths. To illustrate the versatile design of these MSs and the potential of CL in disease diagnosis, these MSs were utilized for in vitro cell targeting and ex vivo CL-excited QD fluorescence (CL-FL) imaging of atherosclerotic plaques in rats. It was shown that by utilizing both QDs and MSs with a high-refractive-index-oil core, the CL emission increases by four-fold at longer wavelengths. Furthermore, we demonstrate that these MSs generate both an in vivo and ex vivo contrast signal. The design concept of utilizing QDs and high-index core MSs may contribute to future developments of in vivo CL imaging.

Interferometric synthetic aperture microscopy (ISAM)

Abstract: The trade-off between transverse resolution and depth-of-field, and the mitigation of optical aberrations, are long-standing problems in optical imaging. The deleterious impact of these problems on three-dimensional tomography increases with numerical aperture (NA), and so they represent a significant impediment for realtime cellular resolution tomography over the typical imaging depths achieved with OCT. With optical coherence microscopy (OCM) [1], which utilizes higher-NA optics than OCT, the depth-of-field is severely reduced [1, 2], and it has been

Handheld Device for Identification of Microbiological Constituents in the Middle Ear

Abstract: Methods and apparatus for identifying microbiological constituents in the middle ear. A spectrometer receives Raman-scattered light from the region of the tympanic membrane and resolves spectral features of the Raman-scattered light. A processor receives the interferometry signal and the Raman signal, and generates a Raman spectrum of the tympanic membrane and material adjacent thereto. In some embodiments of the invention, low-coherence light and substantially monochromatic excitation light are directed onto a tympanic membrane of the ear of a person via an otoscopic tip that abuts the ear canal. An interferometer combines scattered low-coherence light from the ear tissue with a reference signal to generate an interferometric signal.

Enhancement of optical coherence microscopy in turbid media by an optical parametric amplifier.

Abstract: We report the enhancement in imaging performance of a spectral-domain optical coherence microscope (OCM) in turbid media by incorporating an optical parametric amplifier (OPA). The OPA provides a high level of optical gain to the sample arm, thereby improving the signal-to-noise ratio of the OCM by a factor of up to 15 dB. A unique nonlinear confocal gate is automatically formed in the OPA, which enables selective amplification of singly scattered (ballistic) photons against the multiply-scattered light background. Simultaneous enhancement in both imaging depth and spatial resolution in imaging microstructures in highly light-scattering media are demonstrated with the combined OPA-OCM setup. Typical OCM inteferograms (left) and images (right) without and with OPA.

Polarization-sensitive interferometric synthetic aperture microscopy

Abstract: 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.

Retinal imaging with en face and cross-sectional optical coherence tomography delineates outer retinal changes in cancer-associated retinopathy secondary to Merkel cell carcinoma.

Abstract: The study aims to correlate Fourier-domain optical coherence tomography (FD-OCT) with Goldmann visual field (GVF) to show the photoreceptor (PR) structure and function relationship in the first described case of cancer-associated retinopathy (CAR) from Merkel cell carcinoma. A case study of a patient with CAR who was imaged with serial GVF and FD-OCT over a 2-year period was carried out. En face images were created using a custom algorithm from the volumetric Fourier-domain OCT scans at the PR level. The areas of decreased PR reflectivity on the en face images were compared with GVF obtained at the same time point. Regions of reduced signal on en face scans corresponded with the position and shape of the GVF scotomas. Initially, the vision improved without PR changes. Cross-sectional OCTs showed early recovery of the outer nuclear layer and later improvement in the nerve fiber layer. Worsening vision corresponded with recurrence of the underlying cancer. Progressive global retinal atrophy was seen over time. Merkle cell carcinoma can cause CAR. Retinal function recovered without structural PR recovery. Transient vision improvements in treated CAR patients may be due to layers other than the PRs, but eventual vision decline results from significant progressive retinal atrophy.

A computational approach to high-resolution imaging of the living human retina without hardware adaptive optics

Abstract: We demonstrate high-resolution imaging of the living human retina by computationally correcting highorder ocular aberrations. These corrections are performed post-acquisition and without the need for a deformable mirror or wavefront sensor that are commonly employed in hardware adaptive optics (HAO) systems. With the introduction of HAO to ophthalmic imaging, high-resolution near diffraction-limited imaging of the living human retina has become possible. The combination of a deformable mirror, wavefront sensor, and supporting hardware/software, though, can more than double the cost of the underlying imaging modality, in addition to significantly increasing the system complexity and sensitivity to misalignment. Optical coherence tomography (OCT) allows 3-D imaging in addition to naturally providing the complex optical field of backscattered light. This is unlike a scanning laser ophthalmoscope which measures only the intensity of the backscattered light. Previously, our group has demonstrated the utility of a technique called computational adaptive optics (CAO) which utilizes the complex field measured with OCT to computationally correct for optical aberrations in a manner similar to HAO. Until now, CAO has been applied to ex vivo imaging and in vivo skin imaging. Here, we demonstrate in vivo imaging of cone photoreceptors using CAO. Additional practical considerations such as imaging speed, and stability are discussed.

Smart phone attachment for 3-D optical coherence tomography imaging

Abstract: Methods and apparatus for tomographic imaging of a sample. Low-coherence light is split into a sample path and a reference path. A steering optic recombines light in the reference path with light scattered by a sample onto a camera having an areal focal plane array of detector elements such that light in the reference path and light scattered by the sample are characterized by respectively offset propagation vectors at incidence upon the camera. A processor derives depth information from light scattered by the sample on the basis of interference fringes between light in the reference path and light scattered by the sample. The apparatus tracks lateral motion and may be hand-held or attached to a mobile device such as a smartphone, thus enabling 3-D imaging with the mobile device.

Erratum to optical coherence tomography for the diagnosis and evaluation of human otitis media [J Korean Med Sci 2015; 30: 328-335]

Abstract: This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. pISSN 1011-8934 eISSN 1598-6357 http://dx.doi.org/10.3346/jkms.2015.30.6.834 • J Korean Med Sci 2015; 30: 834 CORRESPONDENCE

Biophotonics for assessing breast cancer

Abstract: Biophotonic imaging is rapidly emerging as a tool for clinical assessment of breast cancer at the microscopic and macroscopic scale. Optical biopsy uses near-infrared (NIR) light to analyze tumor margins and lymph nodes with micron-scale resolution intraoperatively. Optical mammography uses NIR light to gain spectroscopic information and three-dimensional images of whole breast tissues noninvasively. The following chapter will review the current research in the application of biophotonic imaging methods such as optical coherence tomography, multiphoton microscopy, vibrational imaging, near-infrared spectroscopy, diffuse optical tomography, and multimodal macroscale imaging for noninvasive diagnosis and intraoperative imaging of breast tumors and lymph nodes.

Computational adaptive optics for broadband optical interferometric tomography of biological tissue

Abstract: High-resolution real-time tomography of biological tissues is important for many areas of biological investigations and medical applications. Cellular level optical tomography, however, has been challenging because of the compromise between transverse imaging resolution and depth-of-field, the system and sample aberrations that may be present, and the low imaging sensitivity deep in scattering tissues. The use of computed optical imaging techniques has the potential to address several of these long-standing limitations and challenges. Two related techniques are interferometric synthetic aperture microscopy (ISAM) and computational adaptive optics (CAO). Through three-dimensional Fourierdomain resampling, in combination with high-speed OCT, ISAM can be used to achieve high-resolution in vivo tomography with enhanced depth sensitivity over a depth-of-field extended by more than an order-of-magnitude, in realtime. Subsequently, aberration correction with CAO can be performed in a tomogram, rather than to the optical beam of a broadband optical interferometry system. Based on principles of Fourier optics, aberration correction with CAO is performed on a virtual pupil using Zernike polynomials, offering the potential to augment or even replace the more complicated and expensive adaptive optics hardware with algorithms implemented on a standard desktop computer. Interferometric tomographic reconstructions are characterized with tissue phantoms containing sub-resolution scattering particles, and in both ex vivo and in vivo biological tissue. This review will collectively establish the foundation for high-speed volumetric cellular-level optical interferometric tomography in living tissues.

Automation of interferometric synthetic aperture microscopy

Abstract: In this paper, we present an automated algorithm framework for determining the optimal parameters for interferometric synthetic aperture microscopy (ISAM). Three stages of ISAM reconstruction, including dispersion correction, spectral domain resampling and computational adaptive optics (CAO) aberration correction are automated. This algorithm framework significantly lowers the background requirement for operating and calibrating ISAM machines, while achieving fast, near-optimal ISAM reconstruction on optical coherence tomography (OCT) datasets.

Finding bugs in your ear: Clinical imaging of middle-ear infections and biofilms using OCT

Abstract: Handheld OCT scanners have been developed for screening and characterizing middle-ear infections that are highly prevalent in the pediatric population. OCT enables non-invasive identification of bacterial biofilms that will impact clinical treatment of this disease.

Optical coherence microscopy using hardware and computational adaptive optics

Abstract: Aberrations can be compensated for by adaptive optics. In this paper, we combined adaptive optics with optical coherence microscopy for high resolution imaging. Comparisons between hardware and computational adaptive optics are presented.