In vivo measurement of the biomechanical properties of human skin with motion-corrected Brillouin microscopy.

Biomechanical testing of human skin in vivo is important to study the aging process and pathological conditions such as skin cancer. Brillouin microscopy allows the all-optical, non-contact visualization of the mechanical properties of cells and tissues over space. Here, we use the combination of Brillouin microscopy and optical coherence tomography for motion-corrected, depth-resolved biomechanical testing of human skin in vivo. We obtained two peaks in the Brillouin spectra for the epidermis, the first at 7 GHz and the second near 9-10 GHz. The experimentally measured Brillouin frequency shift of the dermis is lower compared to the epidermis and is 6.8 GHz, indicating the lower stiffness of the dermis.

Motion-artifact-free single shot two-beam optical coherence elastography system.

Significance: The assessment of the biomechanical properties of the skin using various imaging techniques has been used as a diagnostic tool in dermatology. Optical coherence elastography (OCE) is one of the techniques that allows for the measurement of elastic properties. OCE relies on measuring tissue displacements induced by external sources. Measuring the tissue's mechanical properties in vivo using OCE is often challenging due to bulk tissue movement. Aim: This study aimed to develop an OCE system that allows for minimizing the effects of bulk tissue movements. To achieve this, we designed a two-beam OCE system that simultaneously measures the tissue displacement at two locations on the sample. This allows for cancelling the effect of the tissue bulk movement, which is common to both measurement points. Approach: We used a piezoelectric transducer to generate surface acoustic waves (SAW) in the sample. The velocity of the excited waves, which is proportional to the rigidity of the sample, was measured by calculating the phase delay of the SAW at two locations on the sample. Simultaneous measurement at two locations was achieved by dividing a single light beam into two by focusing on the sample at two different locations. The two beams travel different optical path lengths, and the reflected signals were depth encoded in a single optical coherence tomography scan using a single reference beam. Results: The system was characterized using different tissue-mimicking phantoms and the skin of healthy volunteers at the wrist and the palm. We achieved an approximately 50-fold increase in phase sensitivity measurement. Conclusions: We designed a simple two-beam OCE system that effectively minimizes the effect of tissue movement. We believe that the presented system will find immediate applications in the clinic to monitor the progression of systemic sclerosis disease.

Small leucine-rich proteoglycans inhibit CNS regeneration by modifying the structural and mechanical properties of the lesion environment.

Extracellular matrix (ECM) deposition after central nervous system (CNS) injury leads to inhibitory scarring in humans and other mammals, whereas it facilitates axon regeneration in the zebrafish. However, the molecular basis of these different fates is not understood. Here, we identify small leucine-rich proteoglycans (SLRPs) as a contributing factor to regeneration failure in mammals. We demonstrate that the SLRPs chondroadherin, fibromodulin, lumican, and prolargin are enriched in rodent and human but not zebrafish CNS lesions. Targeting SLRPs to the zebrafish injury ECM inhibits axon regeneration and functional recovery. Mechanistically, we find that SLRPs confer mechano-structural properties to the lesion environment that are adverse to axon growth. Our study reveals SLRPs as inhibitory ECM factors that impair axon regeneration by modifying tissue mechanics and structure, and identifies their enrichment as a feature of human brain and spinal cord lesions. These findings imply that SLRPs may be targets for therapeutic strategies to promote CNS regeneration.

Deep Tissue Characterization with Optical Coherence Elastography: A Comparison of Different Methods.

The measurement of the biomechanical properties of the skin is of great interest since these properties play an important role in the development of several diseases such as skin cancer and systemic sclerosis. In this direction, several diagnostic tools have been developed to analyze the mechanical properties of the skin. Optical coherence elastography (OCE) is one of the emerging imaging techniques used for the characterization of the mechanical properties of the tissue quantitatively. In systemic sclerosis patients, the measurement of the mechanical properties of the deeper skin layers is desirable compared to the superficial layers. There are several variants of OCE that exist, but it is still not clear which method is more suitable for the measurement of the mechanical properties of the deeper tissue. In this work, we tested three common methods, the pulsed excitation method, the continuous wave excitation method, and the resonant frequency method, for the measurement of the mechanical properties of the deeper layers in the tissue. We found out that the pulsed wave excitation method provides the most reliable measurements in the shortest possible time compared to the other two methods.

Depth of focus extension in optical coherence tomography using ultrahigh chromatic dispersion of zinc selenide.

We report a novel technique to overcome the depth-of-focus limitation in optical coherence tomography (OCT) using chromatic dispersion of zinc selenide lens. OCT is an established method of optical imaging, which found numerous biomedical applications. However, the depth scanning range of high-resolution OCT is limited by its depth of focus. Chromatic dispersion of zinc selenide lens allows to get high lateral resolution along extended depth of focus, because the different spectral components are focused at a different position along axes of light propagation. Test measurements with nanoparticle phantom show 2.8 times extension of the depth of focus compare to the system with a standard achromatic lens. The feasibility of biomedical applications was demonstrated by ex vivo imaging of the pig cornea and chicken fat tissue.

Feasibility and Safety of Tethered Capsule Endomicroscopy in Patients With Barrett's Esophagus in a Multi-Center Study.

BACKGROUND & AIMS: Tethered capsule endomicroscopy (TCE) involves swallowing a small tethered pill that implements optical coherence tomography (OCT) imaging, procuring high resolution images of the whole esophagus. Here, we demonstrate and evaluate the feasibility and safety of TCE and a portable OCT imaging system in patients with Barrett's esophagus (BE) in a multi-center (5-site) clinical study. METHODS: Untreated patients with BE as per endoscopic biopsy diagnosis were eligible to participate in the study. TCE procedures were performed in unsedated patients by either doctors or nurses. After the capsule was swallowed, the device continuously obtained 10-µm-resolution cross-sectional images as it traversed the esophagus. Following imaging, the device was withdrawn through mouth, and disinfected for subsequent reuse. BE lengths were compared to endoscopy findings when available. OCT-TCE images were compared to volumetric laser endomicroscopy (VLE) images from a patient who had undergone VLE on the same day as TCE. RESULTS: 147 patients with BE were enrolled across all sites. 116 swallowed the capsule (79%), 95/114 (83.3%) men and 21/33 (63.6%) women (P = .01). High-quality OCT images were obtained in 104/111 swallowers (93.7%) who completed the procedure. The average imaging duration was 5.55 ± 1.92 minutes. The mean length of esophagus imaged per patient was 21.69 ± 5.90 cm. A blinded comparison of maximum extent of BE measured by OCT-TCE and EGD showed a strong correlation (r = 0.77-0.79). OCT-TCE images were of similar quality to those obtained by OCT-VLE. CONCLUSIONS: The capabilities of TCE to be used across multiple sites, be administered to unsedated patients by either physicians or nurses who are not expert in OCT-TCE, and to rapidly and safely evaluate the microscopic structure of the esophagus make it an emerging tool for screening and surveillance of BE patients. Clinical trial registry website and trial number: NCT02994693 and NCT03459339.

Smartphone-based multimodal tethered capsule endoscopic platform for white-light, narrow-band, and fluorescence/autofluorescence imaging.

Multimodal low-cost endoscopy is highly desirable in poor resource settings such as in developing nations. In this work, we developed a smartphone-based low-cost, reusable tethered capsule endoscopic platform that allows white-light, narrowband, and fluorescence/autofluorescence imaging of the esophagus. The ex-vivo studies of swine esophagus were performed and compared with a commercial endoscope to test the white-light imaging capabilities of the endoscope. The efficacy of the capsule for narrow-band imaging was tested by imaging the vascularization of the tongue. To determine the autofluorescence/fluorescence capability of the endoscope, fluorescein dye with different concentrations was imaged. Furthermore, swine esophagus injected with fluorescein dye was imaged using the fluorescence/autofluorescence and the white-light imaging modules, ex-vivo. The overall cost of the capsules is approximately 12 €, 15 €, and 42 € for the white light imaging, the narrow-band imaging, and the fluorescence/autofluorescence imaging respectively. In addition, the cost of the laser source module required for the narrow-band imaging and the fluorescence/autofluorescence imaging is approximately 218 €. This device will open the possibility of imaging the esophagus in underprivileged areas.

Novel input polarisation independent endoscopic cross-polarised optical coherence tomography probe.

Lead by the original idea to perform noninvasive optical biopsies of various tissues, optical coherence tomography found numerous medical applications within the last two decades. The interference based imaging technique opens the possibility to visualise subcellular morphology up to an imaging depth of 3 mm and up to micron level axial and lateral resolution. The birefringence properties of the tissue are visualised with enhanced contrast using polarisation sensitive or cross-polarised optical coherence tomography (OCT) techniques. Although, it requires strict control over the polarisation states, resulting in several polarisation controlling elements. In this work, we propose a novel input-polarisation independent endoscopic system based on cross-polarised OCT. We tested the feasibility of our approach by measuring the polarisation change from a quarter-wave plate for different rotational angles. Further performance tests reveal a lateral resolution of 30 µm and a sensitivity of 103 dB. Images of the human nail bed and cow muscle tissue demonstrate the potential of the system to measure structural and birefringence properties of the tissue endoscopically.

Paclitaxel Drug-Coated Balloon Angioplasty Suppresses Progression and Inflammation of Experimental Atherosclerosis in Rabbits.

Paclitaxel drug-coated balloons (DCBs) reduce restenosis, but their overall safety has recently raised concerns. This study hypothesized that DCBs could lessen inflammation and reduce plaque progression. Using 25 rabbits with cholesterol feeding- and balloon injury-induced lesions, DCB-percutaneous transluminal angioplasty (PTA), plain PTA, or sham-PTA (balloon insertion without inflation) was investigated using serial intravascular near-infrared fluorescence-optical coherence tomography and serial intravascular ultrasound. In these experiments, DCB-PTA reduced inflammation and plaque burden in nonobstructive lesions compared with PTA or sham-PTA. These findings indicated the potential for DCBs to serve safely as regional anti-atherosclerosis therapy.

Input polarization-independent polarization-sensitive optical coherence tomography using a depolarizer.

Polarization-sensitive optical coherence tomography is gaining attention because of its ability to diagnose certain pathological conditions at an early stage. The majority of polarization-sensitive optical coherence tomography systems require a polarization controller and a polarizer to obtain the optimal polarization state of the light at the sample. Such systems are prone to misalignment since any movement of the optical fiber normally coupled to the light source will change the polarization state of the incident beam. We propose and demonstrate an input polarization-independent polarization-sensitive optical coherence tomography system using a depolarizer that works for any input polarization state of the light source. The change in the optical power at the sample for arbitrary input polarized light for the standard polarization-sensitive optical coherence tomography system was found to be approximately 84% compared to 9% for our proposed method. The developed system was used to measure the retardance and optical axis orientation of a quarter-wave plate and the obtained values matched closely to the expectation. To further demonstrate the capability of measuring the birefringent properties of biological samples, we also imaged the nail bed. We believe that the proposed system is a robust polarization-sensitive optical coherence tomography system and that it will improve the diagnostic capabilities in clinical settings.

Optical coherence tomography-guided laser marking with tethered capsule endomicroscopy in unsedated patients.

Tethered capsule endomicroscopy (TCE) is an emerging screening technology that comprehensively obtains microstructural OCT images of the gastrointestinal (GI) tract in unsedated patients. To advance clinical adoption of this imaging technique, it will be important to validate TCE images with co-localized histology, the current diagnostic gold standard. One method for co-localizing OCT images with histology is image-targeted laser marking, which has previously been implemented using a driveshaft-based, balloon OCT catheter, deployed during endoscopy. In this paper, we present a TCE device that scans and targets the imaging beam using a low-cost stepper motor that is integrated inside the capsule. In combination with a 4-laser-diode, high power 1430/1450 nm marking laser system (800 mW on the sample and 1s pulse duration), this technology generated clearly visible marks, with a spatial targeting accuracy of better than 0.5 mm. A laser safety study was done on swine esophagus ex vivo, showing that these exposure parameters did not alter the submucosa, with a large, 4-5x safety margin. The technology was demonstrated in living human subjects and shown to be effective for co-localizing OCT TCE images to biopsies obtained during subsequent endoscopy.

Imaging the Human Prostate Gland Using 1-µm-Resolution Optical Coherence Tomography.

CONTEXT.­: The accuracy of needle biopsy for the detection of prostate cancer is limited by well-known sampling errors. Thus, there is an unmet need for a microscopic screening tool that can screen large regions of the prostate comprehensively for cancer. Previous prostate imaging by optical coherence tomography (OCT) has had insufficient resolution for imaging cellular features related to prostate cancer. We have recently developed micro-optical coherence tomography (µOCT) that generates depth-resolved tissue images at a high frame rate with an isotropic resolution of 1 µm. OBJECTIVE.­: To demonstrate that optical images obtained with µOCT provide cellular-level contrast in prostate specimens that will enable differentiation and diagnosis of prostate pathologies. DESIGN.­: Fresh prostate specimens obtained from surgical resections were scanned with µOCT ex vivo. Histologic features in the µOCT images were correlated to the corresponding conventional histology. RESULTS.­: Findings indicate that µOCT is capable of resolving many of the architectural and cellular features associated with benign and neoplastic prostate. CONCLUSIONS.­: Because µOCT can be implemented in a small-diameter flexible probe, this study suggests that high-resolution µOCT imaging may be a useful tool for needle-based virtual biopsy of the prostate gland.

In-line optical fiber metallic mirror reflector for monolithic common path optical coherence tomography probes.

BACKGROUND AND OBJECTIVES: Endoscopic optical coherence tomography probes suffer from various artifacts due to dispersion imbalance and polarization mismatch between reference and sample arm light. Such artifacts can be minimized using a common path approach. In this work, we demonstrate a miniaturized common path probe for optical coherence tomography using an inline fiber mirror. MATERIALS AND METHODS: A common path optical fiber probe suitable for performing high-resolution endoscopic optical coherence tomography imaging was developed. To achieve common path functionality, an inline fiber mirror was fabricated using a thin gold layer. A commercially available swept source engine was used to test the designed probe in a cadaver human coronary artery ex vivo. RESULTS: We achieved a sensitivity of 104 dB for this probe using a swept source optical coherence tomography system. To test the probe, images of a cadaver human coronary artery were obtained, demonstrating the quality that is comparable to those obtained by OCT systems with separate reference arms. Additionally, we demonstrate recovery of ranging depth by use of a Michelson interferometer in the detection path. CONCLUSION: We developed a miniaturized monolithic inline fiber mirror-based common path probe for optical coherence tomography. Owing to its simplicity, our design will be helpful in endoscopic applications that require high-resolution probes in a compact form factor while reducing system complexity. Lasers Surg. Med. 50:230-235, 2018. © 2017 Wiley Periodicals, Inc.

Astigmatism corrected common path probe for optical coherence tomography.

BACKGROUND AND OBJECTIVES: Optical coherence tomography (OCT) catheters for intraluminal imaging are subject to various artifacts due to reference-sample arm dispersion imbalances and sample arm beam astigmatism. The goal of this work was to develop a probe that minimizes such artifacts. MATERIALS AND METHODS: Our probe was fabricated using a single mode fiber at the tip of which a glass spacer and graded index objective lens were spliced to achieve the desired focal distance. The signal was reflected using a curved reflector to correct for astigmatism caused by the thin, protective, transparent sheath that surrounds the optics. The probe design was optimized using Zemax, a commercially available optical design software. Common path interferometric operation was achieved using Fresnel reflection from the tip of the focusing graded index objective lens. The performance of the probe was tested using a custom designed spectrometer-based OCT system. RESULTS: The probe achieved an axial resolution of 15.6 µm in air, a lateral resolution 33 µm, and a sensitivity of 103 dB. A scattering tissue phantom was imaged to test the performance of the probe for astigmatism correction. Images of the phantom confirmed that this common-path, astigmatism-corrected OCT imaging probe had minimal artifacts in the axial, and lateral dimensions. CONCLUSIONS: In this work, we developed an astigmatism-corrected, common path probe that minimizes artifacts associated with standard OCT probes. This design may be useful for OCT applications that require high axial and lateral resolutions. Lasers Surg. Med. 49:312-318, 2017. © 2016 Wiley Periodicals, Inc.

In vivo imaging of airway cilia and mucus clearance with micro-optical coherence tomography.

We have designed and fabricated a 4 mm diameter rigid endoscopic probe to obtain high resolution micro-optical coherence tomography (µOCT) images from the tracheal epithelium of living swine. Our common-path fiber-optic probe used gradient-index focusing optics, a selectively coated prism reflector to implement a circular-obscuration apodization for depth-of-focus enhancement, and a common-path reference arm and an ultra-broadbrand supercontinuum laser to achieve high axial resolution. Benchtop characterization demonstrated lateral and axial resolutions of 3.4 µm and 1.7 µm, respectively (in tissue). Mechanical standoff rails flanking the imaging window allowed the epithelial surface to be maintained in focus without disrupting mucus flow. During in vivo imaging, relative motion was mitigated by inflating an airway balloon to hold the standoff rails on the epithelium. Software implemented image stabilization was also implemented during post-processing. The resulting image sequences yielded co-registered quantitative outputs of airway surface liquid and periciliary liquid layer thicknesses, ciliary beat frequency, and mucociliary transport rate, metrics that directly indicate airway epithelial function that have dominated in vitro research in diseases such as cystic fibrosis, but have not been available in vivo.

Ultra-high dynamic range electro-optic sampling for detecting millimeter and sub-millimeter radiation.

Time-domain spectroscopy using coherent millimeter and sub-millimeter radiation (also known as terahertz radiation) is rapidly expanding its application, owing greatly to the remarkable advances in generating and detecting such radiation. However, many current techniques for coherent terahertz detection have limited dynamic range, thus making it difficult to perform some basic experiments that need to directly compare strong and weak terahertz signals. Here, we propose and demonstrate a novel technique based on cross-polarized spectral-domain interferometry to achieve ultra-high dynamic range electro-optic sampling measurement of coherent millimeter and sub-millimeter radiation. In our scheme, we exploit the birefringence in a single-mode polarization maintaining fiber in order to measure the phase change induced by the electric field of terahertz radiation in the detection crystal. With our new technique, we have achieved a dynamic range of 7 × 10(6), which is 4 orders of magnitude higher than conventional electro-optic sampling techniques, while maintaining comparable signal-to-noise ratio. The present technique is foreseen to have great impact on experiments such as linear terahertz spectroscopy of optically thick materials (such as aqueous samples) and nonlinear terahertz spectroscopy, where the higher dynamic range is crucial for proper interpretation of experimentally obtained results.

µOCT imaging using depth of focus extension by self-imaging wavefront division in a common-path fiber optic probe.

Optical coherence tomography (OCT) is an attractive medical modality due to its ability to acquire high-resolution, cross-sectional images inside the body using flexible, small-diameter, scanning fiber optic probes. Conventional, cross-sectional OCT imaging technologies have approximately 10-µm axial resolution and 30-µm lateral resolution, specifications that enable the visualization of microscopic architectural morphology. While this resolution is useful for many clinical applications, it is insufficient for resolving individual cells that characterize many diseases. To address this gap, a supercontinuum-laser-based, µm-resolution OCT (µOCT) system and a 500 µm-diameter, extended depth of focus single fiber optic probe for endoscopic and intravascular imaging were designed and fabricated. At the distal tip of the fiber optic probe, a cylindrical waveguide was used to divide the wavefront to provide multiple circular propagation modes. Once transmitted through a relatively high NA lens (NA >0.1), these modes were projected as multiple coaxial foci (~3 µm full width at half maximum (FWHM)) over a greatly extended focal depth range. The distal tip of the probe also contained a common-path reference reflectance to minimize polarization and dispersion imbalances between sample and reference arm light. Measurements showed that the probe provides a 20-fold depth of focus extension, maintaining a 3-5 µm lateral resolution (FWHM of PSF) and a 2 µm axial resolution over a depth range of approximately 1 mm. These results suggest that this new optical configuration will be useful for achieving high-resolution, cross-sectional OCT imaging in catheter/endoscope-based medical imaging devices.

Characterization of tissue-engineered posterior corneas using second- and third-harmonic generation microscopy.

Three-dimensional tissues, such as the cornea, are now being engineered as substitutes for the rehabilitation of vision in patients with blinding corneal diseases. Engineering of tissues for translational purposes requires a non-invasive monitoring to control the quality of the resulting biomaterial. Unfortunately, most current methods still imply invasive steps, such as fixation and staining, to clearly observe the tissue-engineered cornea, a transparent tissue with weak natural contrast. Second- and third-harmonic generation imaging are well known to provide high-contrast, high spatial resolution images of such tissues, by taking advantage of the endogenous contrast agents of the tissue itself. In this article, we imaged tissue-engineered corneal substitutes using both harmonic microscopy and classic histopathology techniques. We demonstrate that second- and third-harmonic imaging can non-invasively provide important information regarding the quality and the integrity of these partial-thickness posterior corneal substitutes (observation of collagen network, fibroblasts and endothelial cells). These two nonlinear imaging modalities offer the new opportunity of monitoring the engineered corneas during the entire process of production.

Preparing uniform-thickness corneal endothelial grafts from donor tissues using a non-amplified femtosecond laser.

Corneal grafts for Descemet's Stripping Automated Endothelial Keratoplasty are commonly prepared using mechanical microkeratomes. However, the cuts produced in such way render corneal lenticules that are thinner centrally than peripherally, thus inducing a hyperopic shift. Here we describe a novel device for preparing donor corneal grafts, in which a single low-energy femtosecond laser system is used as both a light source for optical coherence tomography and for cutting the graft illuminating from the endothelial side. The same laser is first utilized to obtain three-dimensional optical coherence tomography images of the donor tissue for guiding the dissection and obtaining grafts of uniform thickness with no applanation or contact. This device allows an optimal procedure for preparing consistently thin posterior grafts for transplantation.

Self-referenced spectral domain interferometry for improved signal-to-noise measurement of terahertz radiation.

In this work, we demonstrate self-referenced spectral domain interferometry for the electro-optic sampling of terahertz (THz) electric fields. This technique allows reduction of the phase noise of the measurement, thus increasing the signal-to-noise ratio (SNR). Using the proposed technique, we achieve a more than sixfold improvement in the SNR of the detected THz electric field.