Biomechanical testing of human skin in vivo is important for studying the aging process and pathological conditions such as skin cancer. Brillouin microscopy allows non-contact measurements of the bulk elastic modulus of the skin, providing information on the mechanical properties of the tissue. Here, we use the combination of Brillouin microscopy and OCT imaging for motion-corrected, depth-resolved biomechanical testing of human skin in vivo. We found that the experimentally measured Brillouin frequency shift of the epidermis is higher than that of the dermis, indicating the higher stiffness of the upper skin layer.
The analysis of the biomechanical properties is a key factor in disease diagnosis at an earlier stage. It has found several applications in fields such as systemic sclerosis. Optical coherence elastography (OCE) is one of the techniques which has been used to characterize the mechanical properties of the skin. We designed a two-beam OCE system with better phase stability (50 times) than a single-point measurement. This system allows for minimizing bulk tissue movement. We used a piezo transducer to excite acoustic waves in the tissue and two beams to measure the acoustic wave velocity. We measured the elasticity of different ager phantoms and skin.
SignificanceThe 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.AimThis 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.ApproachWe 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.ResultsThe 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.ConclusionsWe 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.
We developed a novel fiber-based minimalistic system for cross-polarization optical coherence tomography using polarization-maintaining fiber in the sample arm. A long section of polarization-maintaining fiber was placed in the sample arm which acted as a delay line. Co- and cross-polarized images appear at different depth positions within the image. Test measurements with a quarter-wave plate demonstrate the sensitivity of the system to the birefringence of the sample. The signal in the images is not sensitive to the bending of polarization maintaining fiber in the sample arm, making the system a promising tool for endoscopy and clinical applications.
Tethered capsule endomicroscopy (TCE) is a recently developed form of in vivo microscopy based on optical coherence tomography (OCT). With TCE, a small tethered pill is swallowed, procuring high resolution microscopic images of the esophageal wall. TCE does not require sedation and is thus a more rapid and convenient procedure comparing to traditional endoscopic examination. Our group and others have successfully conducted OCT-TCE in pilot, single-center studies that demonstrated the potential of this technology for upper GI tract diagnosis. Here, we demonstrate and evaluate the feasibility and safety of a next generation OCT-TCE system and device in patients with Barrett’s esophagus (BE) and report the initial longitudinal analysis of the natural history of BE.
Multimodal low-cost endoscopy is desirable in poor resource settings. Here, we developed smartphone-based low-cost, reusable tethered capsule endoscope that allows white-light, narrowband, and fluorescence/autofluorescence imaging of esophagus
Hyperspectral-imaging is label-free imaging technique. We designed hyperspectral source based on chromatic dispersion property of off-the-shelf lenses, that can be incorporated into standard endoscope/microscope to perform hyperspectral-imaging.
While the most common method used to evaluate and survey patients with Barrett’s Esophagus (BE) is endoscopic biopsy, this procedure is invasive, time-consuming, and suffers from sampling errors. Moreover, it requires patient sedation that increases cost and mandates its operation in specialized settings. Our lab has developed a new imaging tool termed tethered capsule endomicroscopy (TCE) that involves swallowing a tethered capsule which utilizes optical coherence tomography (OCT) to obtain three-dimensional microscopic (10µm) images of the entire esophageal wall as it traverses the luminal organ via peristalsis or is retrieved by pulling up tether. As opposed to endoscopy, TCE procedure is non-invasive, doesn’t require patient sedation and mitigates sampling error by evaluating the microscopic structure of the entire esophagus. The merits of TCE make it a suitable device to investigate the microscopic natural history of BE in a longitudinal manner.
Here, we present our initial experience of a multicenter (5-site) clinical trial to study the microscopic natural history of BE. The TCE device used for the study is the new generation capsule with the ball lens optical configuration and a distal scan stepper motor, which provides 30µm (lateral) resolution and 40Hz imaging rate. The portable OCT imaging system is a custom in-house built swept source system and provides 7µm (axial) at a 100 kHz A-line rate with a center wavelength of ~1310 nm. To date, we have successfully enrolled 69 subjects at all sites (MGH: 33, Columbia University: 11, Kansas City VA: 10, Mayo Jacksonville: 8, Mayo Rochester: 7) and 59 have swallowed the capsule (85.5%). There have been no reported adverse events associated with TCE procedure. High-quality OCT images were reliably obtained from patients who swallowed the device, and BE tissues were identified by expert readers. Our initial experience with TCE in a multicenter study demonstrates that this technology is easy to use and efficient in multiple clinical settings. Completion of this longitudinal study is likely to provide new insights on the temporal progression of BE that may impact management strategies.
Background: Birefringent crystals such as cholesterol and monosodium urate have recently been identified as possible pharmacologic targets for the treatment of coronary artery disease. The size of these crystals can be very small (on the order of 1 µm), making them difficult to identify. To image these microscopic crystals and enhance contrast, we modified existing micro optical coherence tomography system so that it was capable of obtaining polarization-sensitive images (PS-µOCT).
A spectrometer-based PS-µOCT system was developed using a 270 nm wide broadband light source centered at 765 nm. Light was polarized using a polarizer and coupled to a SMF. The polarized light after SMF was divided into reference and sample arms using a beam splitter. Images of orthogonal polarization states were acquired sequentially by inserting and removing a quarter wave plate in the reference arm. The orthogonal PS- µOCT image components were used to generate birefringent images of the tissue.
The axial resolution of the PS-µOCT system was 1.9 mm and the lateral resolution was 2 microns and the SNR was 92 dB. PS-µOCT was able to clearly identify isolated cholesterol and uric acid crystals. When used to image cadaver coronary arteries, the PS-µOCT images of crystals had up to 11 dB improved contrast compared to images obtained with a standard µOCT system.
Results show that the use of PS-µOCT improves image contrast for isolated crystals and crystals within coronary atherosclerotic plaque and suggest that it could be useful for understanding their roles in the development and progression of coronary artery disease.
Tethered capsule endomicroscopy (TCE) is a new method for performing comprehensive microstructural OCT imaging of gastrointestinal (GI) tract in unsedated patients in a well-tolerated and cost-effective manner. These features of TCE bestow it with significant potential to improve the screening, surveillance and management of various upper gastrointestinal diseases. To achieve clinical adoption of this imaging technique, it is important to validate it with co-registered histology, the current diagnostic gold standard. One such method for co-registering OCT images with histology is laser cautery marking, previously demonstrated using a balloon-centering OCT catheter that operates in conjunction with sedated endoscopy. With laser marking, an OCT area of interest is identified on the screen and this target is marked in the patient by exposing adjacent tissue to laser light that is absorbed by water, creating superficial, visible marks on the mucosal surface. Endoscopy can then be performed after the device is removed and biopsies taken from the marks. In this talk, we will present the design of a tethered capsule laser marking device that uses a distal stepper motor to perform high precision (< 0.5 mm accuracy) laser targeting and high quality OCT imaging. Ex vivo animal tissue tests and pilot human clinical studies using this technology will be presented.
In optical coherence tomography (OCT) systems, axial resolution improves with increasing light source bandwidth. However, dispersion imbalance between the sample and reference arms can degrade axial resolution and signal to noise ratio, a significant issue for ultrahigh-resolution OCT systems. In this work, we demonstrate a novel technique for estimating and compensating for OCT system dispersion, that is unique from previously reported methods in that it compensates all orders of system dispersion. Dispersion phase was estimated by first measuring the phase from of the spectrogram at two different, reference-sample arm optical path length differences (OPLD) around zero OPLD and then subtracting the two phase values to obtain the dispersion phase. This phase can be used to compensate the dispersion term in the spectrum by multiplying the interference pattern with where k is the wave-vector. This method was tested to compensate the dispersion caused by a 3-mm fused silica window in one arm of an ultrahigh spectral domain OCT system in our laboratory that utilizes a light source with a 850 nm center wavelength, 300 nm bandwidth. Using our dispersion compensation technique, the experimentally measured axial resolution of the system was fully recovered to match the theoretical resolution, improving from 10.6µm to 1.85µm in air. These results suggest that this dispersion compensation method may be useful to avoid axial resolution degradation due to dispersion effects in ultrahigh-resolution OCT systems that employ extremely broad band light sources.
Sensorineural hearing loss (SNHL) is the most common sensory deficit in the world, caused by damage to cellular structures within the inner ear, or cochlea. Visualization of the cellular pathology underlying different types of SNHL has been difficult due to the small size of the cochlea, its complex three-dimensional structure, and embedded location within the temporal bone. Micro-optical coherence tomography (µOCT) is a recently-developed cross-sectional imaging technology that can obtain images with sufficient detail to elucidate specific aetiologies of SNHL. In this work, we developed a high resolution, ultra-small-diameter, flexible probe for imaging the human cochlea in situ. The 500 µm diameter, circumferential scanning µOCT imaging probe contains self-imaging wavefront division optics that provide maximal lateral resolution of 2.5 µm and better than 5 µm resolution over an extended depth of focus of 1 mm in air. Using a supercontinuum light source with a 300 nm bandwidth and a common path interferometery configuration, axial resolution is 1.9 µm in air. Images of 3D-intact cochleae extracted from human cadavers were acquired with the µOCT probe in situ; these images demonstrate the system’s ability to visualize the entire cross-section of the scala tympani, in addition to cellular structures in the cochlea’s sensory epithelium, the organ of Corti and bundles of auditory nerves. These results suggest that this new device has the potential to facilitate personalized diagnosis and therapy for SNHL.
Background: Uric acid crystals have recently been identified as a possible therapeutic target for coronary artery disease. Being subcellular in size, it is difficult to identify these crystals in situ. Micro optical coherence tomography (Micro-OCT) allows one to image subcellular structures with 1–micron resolution. Even though Micro-OCT should be capable of resolving urate crystals, it’s difficult to differentiate these structures from other scattering particles within tissue. In this work we developed a novel polarization sensitive micro OCT (ps-Micro-OCT) system for identification of uric acid crystals.
Methods: A spectrometer based ps-Micro-OCT system was developed using a broadband light source. The broadband input light was divided into reference and sample signals using a beam splitter. The reference signal was further divided into two polarized signals with different polarization states. Reflected reference and sample signals were combined and sent to a spectrometer that recorded the interference signal.
Results: To test the performance of system, a mirror was used as sample and a quarter wave-plate was placed in the sample path. The measured quarter wave-plate angle values matched closely to actual angle values. Next we prepared uric acid crystals in our lab and imaged them using this system.We were able to image and identify these crystals based on polarization measurements.
Conclusion: In this work we imaged and identified uric acid crystals using a newly developed ps-Micro-OCT system. The proposed technique will enable imaging uric acid crystals in coronary artery.
Background: Dispersion imbalance and polarization mismatch between the reference and sample arm signals can lead to image quality degradation in optical coherence tomography (OCT). One approach to reduce these image artifacts is to employ a common-path geometry in fiber-based probes. In this work, we report an 800 um diameter all-fiber common-path monolithic probe for coronary artery imaging where the reference signal is generated using an inline fiber partial reflector.
Methods: Our common-path probe was designed for swept-source based Fourier domain OCT at 1310 nm wavelength. A face of a coreless fiber was coated with gold and spliced to a standard SMF-28 single mode fiber creating an inline partial reflector, which acted as a reference surface. The other face of the coreless fiber was shaped into a ball lens for focusing. The optical elements were assembled within a 560 µm diameter drive shaft, which was attached to a rotary junction. The drive shaft was placed inside a transparent sheath having an outer diameter of 800 µm.
Results: With a source input power of 30mW, the inline common-path probe achieved a sensitivity of 104 dB. Images of human finger skin showed the characteristic layers of skin as well as features such as sweat ducts. Images of coronary arteries ex vivo obtained with this probe enabled visualization of the characteristic architectural morphology of the normal artery wall and known features of atherosclerotic plaque.
Conclusion: In this work, we have demonstrated a common path OCT probe for cardiovascular imaging. The probe is easy to fabricate, will reduce system complexity and overall cost. We believe that this design will be helpful in endoscopic applications that require high resolution and a compact form factor.
Endoscopy, the current standard of care for the diagnosis of upper gastrointestinal (GI) diseases, is not ideal as a screening tool because it is costly, necessitates a team of medically trained personnel, and typically requires that the patient be sedated. Endoscopy is also a superficial macroscopic imaging modality and therefore is unable to provide detailed information on subsurface microscopic structure that is required to render a precise tissue diagnosis. We have overcome these limitations through the development of an optical coherence tomography tethered capsule endomicroscopy (OCT-TCE) imaging device. The OCT-TCE device has a pill-like form factor with an optically clear wall to allow the contained opto-mechanical components to scan the OCT beam along the circumference of the esophagus. Once swallowed, the OCT-TCE device traverses the esophagus naturally via peristalsis and multiple cross-sectional OCT images are obtained at 30-40 μm lateral resolution by 7 μm axial resolution. While this spatial resolution enables differentiation of squamous vs columnar mucosa, crucial microstructural features such as goblet cells (~10 μm), which signify intestinal metaplasia in BE, and enlarged nuclei that are indicative of dysplasia cannot be resolved with the current OCT-TCE technology.
In this work we demonstrate a novel design of a high lateral resolution OCT-TCE device with an extended depth of focus (EDOF). The EDOF is created by use of self-imaging wavefront division multiplexing that produces multiple focused modes at different depths into the sample. The overall size of the EDOF TCE is similar to that of the previous OCT-TCE device (~ 11 mm by 26 mm) but with a lateral resolution of ~ 8 μm over a depth range of ~ 2 mm. Preliminary esophageal and intestinal imaging using these EDOF optics demonstrates an improvement in the ability to resolve tissue morphology including individual glands and cells. These results suggest that the use of EDOF optics may be a promising avenue for increasing the accuracy of OCT-TCE for the diagnosis of upper GI diseases.
Mucociliary clearance (MCC) plays a significant role in maintaining the health of human respiratory system by eliminating foreign particles trapped within mucus. Failure of this mechanism in diseases such as cystic fibrosis and chronic obstructive pulmonary disease (COPD) leads to airway blockage and lung infection, causing morbidity and mortality. The volume of airway mucus and the periciliary liquid encapsulating the cilia, in addition to ciliary beat frequency and velocity of mucociliary transport, are vital parameters of airway health. However, the diagnosis of disease pathogenesis and advances of novel therapeutics are hindered by the lack of tools for visualization of ciliary function in vivo.
Our laboratory has previously developed a 1-µm resolution optical coherence tomography method, termed Micro-OCT, which is capable of visualizing mucociliary transport and quantitatively capturing epithelial functional metrics. We have also miniaturized Micro-OCT optics in a first-generation rigid 4mm Micro-OCT endoscope utilizing a common-path design and an apodizing prism configuration to produce an annular profile sample beam, and reported the first in vivo visualization of mucociliary transport in swine. We now demonstrate a flexible 2.5 mm Micro-OCT probe that can be inserted through the instrument channel of standard flexible bronchoscopes, allowing bronchoscopic navigation to smaller airways and greatly improving clinical utility. Longitudinal scanning over a field of view of more than 400 µm at a frame rate of 40 Hz was accomplished with a driveshaft transduced by a piezo-electric stack motor.
We present characterization and imaging results from the flexible micro-OCT probe and progress towards clinical translation. The ability of the bronchoscope-compatible micro-OCT probe to image mucus clearance and epithelial function will enable studies of cystic fibrosis pathogenesis in small airways, provide diagnosis of mucociliary clearance disorders, and allow individual responses to treatments to be monitored.
Background: Common path probes are highly desirable for optical coherence tomography (OCT) as they reduce system complexity and cost. In this work we report an all-fiber common path side viewing monolithic probe for coronary artery imaging.
Methods: Our common path probe was designed for spectrometer based Fourier domain OCT at 1310 nm wavelength. Light from the fiber expands in the coreless fiber region and then focussed by the ball lens. Reflection from ball lens-air interface served as reference signal. The monolithic ball lens probe was assembled within a 560 µmouter diameter drive shaft which was attached to a rotary junction. The drive shaft was placed inside an outer, transparent sheath of 800 µm diameter.
Results: With a source input power of 25 mW, we could achieve sensitivity of 100.5 dB. The axial resolution of the system was found to be 15.6 µm in air and the lateral resolution (full width half maximum) was approximately 49 µm. As proof of principal, images of skin acquired using this probe demonstrated clear visualization of the stratum corneum, epidermis, and papillary dermis, along with sweat ducts.
Conclusion: In this work we have demonstrated a monolithic, ball lens common, path probe for OCT imaging. The designed ball lens probe is easy to fabricate using a laser splicer. Based on the features and capability of common path probes to provide a simpler solution for OCT, we believe that this development will be an important enhancement for certain types of catheters.
We demonstrate high spatial resolution imaging of a stromal cut in the ex-vivo pig
cornea, using second- and third-harmonic generation microscopy. From these images,
we see in detail how the cut affects the corneal layers. In the beginning of the cut, the
anterior layers, in which the blade is passing through, are disorganized, which could
explain the shadows observed on the images. In the stroma, the cut can be imaged by
third harmonic microscopy, probably due to the χ3 contrast. Although the current
results were obtained from the healthy ex-vivo cornea, it already allows one to
understand the effects of the cut on the tissue characteristics (such as scattering).
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