Multi-MHz MEMS-VCSEL swept-source optical coherence tomography for endoscopic structural and angiographic imaging with miniaturized brushless motor probes.

Swept source optical coherence tomography (SS-OCT) enables volumetric imaging of subsurface structure. However, applications requiring wide fields of view (FOV), rapid imaging, and higher resolutions have been challenging because multi-MHz axial scan (A-scan) rates are needed. We describe a microelectromechanical systems vertical cavity surface-emitting laser (MEMS-VCSEL) SS-OCT technology for A-scan rates of 2.4 and 3.0 MHz. Sweep to sweep calibration and resampling are performed using dual channel acquisition of the OCT signal and a Mach Zehnder interferometer signal, overcoming inherent optical clock limitations and enabling higher performance. We demonstrate ultrahigh speed structural SS-OCT and OCT angiography (OCTA) imaging of the swine gastrointestinal tract using a suite of miniaturized brushless motor probes, including a 3.2 mm diameter micromotor OCT catheter, a 12 mm diameter tethered OCT capsule, and a 12 mm diameter widefield OCTA probe. MEMS-VCSELs promise to enable ultrahigh speed SS-OCT with a scalable, low cost, and manufacturable technology, suitable for a diverse range of imaging applications.

Dual-Gas Quartz-Enhanced Photoacoustic Sensor for Simultaneous Detection of Methane/Nitrous Oxide and Water Vapor.

The development of a dual-gas quartz-enhanced photoacoustic (QEPAS) sensor capable of simultaneous detection of water vapor and alternatively methane or nitrous oxide is reported. A diode laser and a quantum cascade laser (QCL) excited independently and simultaneously both the fundamental and the first overtone flexural mode of the quartz tuning fork (QTF), respectively. The diode laser targeted a water absorption line located at 7181.16 cm-1 (1.392 µm), while the QCL emission wavelength is centered at 7.71 µm and was tuned to target two strong absorption lines of methane and nitrous oxide, located at 1297.47 and 1297.05 cm-1, respectively. Two sets of microresonator tubes were positioned, respectively, at the antinode points of the fundamental and the first overtone flexural modes of the QTF to enhance the QEPAS signal-to-noise ratio. Detection limits of 18 ppb for methane, 5 ppb for nitrous oxide and 20 ppm for water vapor have been achieved at a lock-in integration time of 100 ms.

Tuning forks with optimized geometries for quartz-enhanced photoacoustic spectroscopy.

We report on the design, realization, and performance of novel quartz tuning forks (QTFs) optimized for quartz-enhanced photoacoustic spectroscopy (QEPAS). Starting from a QTF geometry designed to provide a fundamental flexural in-plane vibrational mode resonance frequency of ~16 kHz, with a quality factor of 15,000 at atmospheric pressure, two novel geometries have been realized: a QTF with T-shaped prongs and a QTF with prongs having rectangular grooves carved on both surface sides. The QTF with grooves showed the lowest electrical resistance, while the T-shaped prongs QTF provided the best photoacoustic response in terms of signal-to-noise ratio (SNR). When acoustically coupled with a pair of micro-resonator tubes, the T-shaped QTF provides a SNR enhancement of a factor of 60 with respect to the bare QTF, which represents a record value for mid-infrared QEPAS sensing.

Loss Mechanisms Determining the Quality Factors in Quartz Tuning Forks Vibrating at the Fundamental and First Overtone Modes.

Quartz tuning forks (QTFs) are piezo-transducers that have been implemented for numerous applications, such as chemical gas sensing, atomic force microscopy, rheology, and industrial process control. The most important parameter for QTFs' sensing application is the resonance quality factor (Q-factor). An experimental investigation and theoretical analysis of the influence of QTFs' geometries on the Q-factor of the flexural fundamental and first overtone resonance modes are reported. The resonance frequencies and related Q-factors for five different QTFs have been measured. The QTF response was recorded at different air pressures to investigate the influence of the surrounding medium on the Q-factor. A data analysis demonstrated that air viscous damping is the dominant energy dissipation mechanism for both flexural modes. Thermoelastic and support losses are additional contributions that depend on the QTF geometry. A study of the QTF damping mechanism dependence upon the prong geometry is also provided.

Octupole electrode pattern for tuning forks vibrating at the first overtone mode in quartz-enhanced photoacoustic spectroscopy.

The design, realization, and performance analysis of an octupole electrode pattern configuration intended for the optimization of the charge collection efficiency in quartz tuning forks (QTFs) vibrating at the first overtone in-plane flexural mode is reported. Two QTFs having the same geometry, but differing in the electrode pattern deposited on the QTF prongs, have been realized in order to study the influence of the electrode pattern on the resonance quality factor and electrical resistance. A standard quadrupole pattern (optimized for the fundamental mode) and an octupole electrode layout have been implemented. Although both QTFs show the same resonance quality factor for the first overtone, the octupole pattern provides a reduction of the QTF electrical resistance by more than four times. The sensing performance of the two QTFs has been compared by employing them in a mid-IR quartz-enhanced photoacoustic sensor (QEPAS) system targeting a water absorption line. When operating at the first overtone mode, the QTF with an octupole electrode pattern provides a QEPAS signal more than two times higher with respect to the QTF employing the standard quadrupole configuration.

Cycloid scanning for wide field optical coherence tomography endomicroscopy and angiography in vivo.

Devices that perform wide field-of-view (FOV) precision optical scanning are important for endoscopic assessment and diagnosis of luminal organ disease such as in gastroenterology. Optical scanning for in vivo endoscopic imaging has traditionally relied on one or more proximal mechanical actuators, limiting scan accuracy and imaging speed. There is a need for rapid and precise two-dimensional (2D) microscanning technologies to enable the translation of benchtop scanning microscopies to in vivo endoscopic imaging. We demonstrate a new cycloid scanner in a tethered capsule for ultrahigh speed, side-viewing optical coherence tomography (OCT) endomicroscopy in vivo. The cycloid capsule incorporates two scanners: a piezoelectrically actuated resonant fiber scanner to perform a precision, small FOV, fast scan and a micromotor scanner to perform a wide FOV, slow scan. Together these scanners distally scan the beam circumferentially in a 2D cycloid pattern, generating an unwrapped 1 mm × 38 mm strip FOV. Sequential strip volumes can be acquired with proximal pullback to image centimeter-long regions. Using ultrahigh speed 1.3 µm wavelength swept-source OCT at a 1.17 MHz axial scan rate, we imaged the human rectum at 3 volumes/s. Each OCT strip volume had 166 × 2322 axial scans with 8.5 µm axial and 30 µm transverse resolution. We further demonstrate OCT angiography at 0.5 volumes/s, producing volumetric images of vasculature. In addition to OCT applications, cycloid scanning promises to enable precision 2D optical scanning for other imaging modalities, including fluorescence confocal and nonlinear microscopy.

Microscope-Integrated Intraoperative Ultrahigh-Speed Swept-Source Optical Coherence Tomography for Widefield Retinal and Anterior Segment Imaging.

BACKGROUND AND OBJECTIVE: To demonstrate the feasibility of retinal and anterior segment intraoperative widefield imaging using an ultrahigh-speed, swept-source optical coherence tomography (SS-OCT) surgical microscope attachment. PATIENTS AND METHODS: A prototype post-objective SS-OCT using a 1,050-nm wavelength, 400 kHz A-scan rate, vertical cavity surface-emitting laser (VCSEL) light source was integrated to a commercial ophthalmic surgical microscope after the objective. Each widefield OCT data set was acquired in 3 seconds (1,000 × 1,000 A-scans, 12 × 12 mm2 for retina and 10 × 10 mm2 for anterior segment). RESULTS: Intraoperative SS-OCT was performed in 20 eyes of 20 patients. In six of seven membrane peels and five of seven rhegmatogenous retinal detachment repair surgeries, widefield retinal imaging enabled evaluation pre- and postoperatively. In all seven cataract cases, anterior imaging evaluated the integrity of the posterior lens capsule. CONCLUSIONS: Ultrahigh-speed SS-OCT enables widefield intraoperative viewing in the posterior and anterior eye. Widefield imaging visualizes ocular structures and pathology without requiring OCT realignment. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:94-102.].

Circumferential optical coherence tomography angiography imaging of the swine esophagus using a micromotor balloon catheter.

We demonstrate a micromotor balloon imaging catheter for ultrahigh speed endoscopic optical coherence tomography (OCT) which provides wide area, circumferential structural and angiographic imaging of the esophagus without contrast agents. Using a 1310 nm MEMS tunable wavelength swept VCSEL light source, the system has a 1.2 MHz A-scan rate and ~8.5 µm axial resolution in tissue. The micromotor balloon catheter enables circumferential imaging of the esophagus at 240 frames per second (fps) with a ~30 µm (FWHM) spot size. Volumetric imaging is achieved by proximal pullback of the micromotor assembly within the balloon at 1.5 mm/sec. Volumetric data consisting of 4200 circumferential images of 5,000 A-scans each over a 2.6 cm length, covering a ~13 cm(2) area is acquired in <18 seconds. A non-rigid image registration algorithm is used to suppress motion artifacts from non-uniform rotational distortion (NURD), cardiac motion or respiration. En face OCT images at various depths can be generated. OCT angiography (OCTA) is computed using intensity decorrelation between sequential pairs of circumferential scans and enables three-dimensional visualization of vasculature. Wide area volumetric OCT and OCTA imaging of the swine esophagus in vivo is demonstrated.

Applicability, usability, and limitations of murine embryonic imaging with optical coherence tomography and optical projection tomography.

We present an analysis of imaging murine embryos at various embryonic developmental stages (embryonic day 9.5, 11.5, and 13.5) by optical coherence tomography (OCT) and optical projection tomography (OPT). We demonstrate that while OCT was capable of rapid high-resolution live 3D imaging, its limited penetration depth prevented visualization of deeper structures, particularly in later stage embryos. In contrast, OPT was able to image the whole embryos, but could not be used in vivo because the embryos must be fixed and cleared. Moreover, the fixation process significantly altered the embryo morphology, which was quantified by the volume of the eye-globes before and after fixation. All of these factors should be weighed when determining which imaging modality one should use to achieve particular goals of a study.

Cubic meter volume optical coherence tomography.

Optical coherence tomography (OCT) is a powerful three-dimensional (3D) imaging modality with micrometer-scale axial resolution and up to multi-GigaVoxel/s imaging speed. However, the imaging range of high-speed OCT has been limited. Here, we report 3D OCT over cubic meter volumes using a long coherence length, 1310 nm vertical-cavity surface-emitting laser and silicon photonic integrated circuit dual-quadrature receiver technology combined with enhanced signal processing. We achieved 15 µm depth resolution for tomographic imaging at a 100 kHz axial scan rate over a 1.5 m range. We show 3D macroscopic imaging examples of a human mannequin, bicycle, machine shop gauge blocks, and a human skull/brain model. High-bandwidth, meter-range OCT demonstrates new capabilities that promise to enable a wide range of biomedical, scientific, industrial, and research applications.

Mid-infrared supercontinuum generation spanning 1.8 octaves using step-index indium fluoride fiber pumped by a femtosecond fiber laser near 2 µm.

A nearly two-octave wide coherent mid-infrared supercontinuum is demonstrated in a dispersion-engineered step-index indium fluoride fiber pumped near 2 µm. The pump source is an all-fiber femtosecond laser with 100 fs pulse width, 570 mW average power and 50 MHz repetition rate. The supercontinuum spectrum spans from 1.25 µm to 4.6 µm. Numerical modelling of the supercontinuum spectra show good agreement with the measurements. The coherence of the supercontinuum is calculated using a numerical model and shows a high degree of coherence across the generated bandwidth allowing it to be used for frequency comb applications.

Wideband Electrically-Pumped 1050 nm MEMS-Tunable VCSEL for Ophthalmic Imaging.

In this paper, we present a 1050 nm electrically-pumped micro-electro-mechanically-tunable vertical-cavity-surface-emitting-laser (MEMS-VCSEL) with a record dynamic tuning bandwidth of 63.8 nm, suitable for swept source optical coherence tomography (SS-OCT) imaging. These devices provide reduced cost & complexity relative to previously demonstrated optically pumped devices by obviating the need for a pump laser and associated hardware. We demonstrate ophthalmic SS-OCT imaging with the electrically-pumped MEMS-VCSEL at a 400 kHz axial scan rate for wide field imaging of the in vivo human retina over a 12 mm × 12 mm field and for OCT angiography of the macula over 6 mm × 6 mm & 3 mm × 3 mm fields to show retinal vasculature and capillary structure near the fovea. These results demonstrate the feasibility of electrically pumped MEMS-VCSELs in ophthalmic instrumentation, the largest clinical application of OCT. In addition, we estimate that the 3 dB coherence length in air is 225 meters ± 51 meters, far greater than required for ophthalmic SS-OCT and suggestive of other distance ranging applications.

Ultrahigh speed en face OCT capsule for endoscopic imaging.

Depth resolved and en face OCT visualization in vivo may have important clinical applications in endoscopy. We demonstrate a high speed, two-dimensional (2D) distal scanning capsule with a micromotor for fast rotary scanning and a pneumatic actuator for precision longitudinal scanning. Longitudinal position measurement and image registration were performed by optical tracking of the pneumatic scanner. The 2D scanning device enables high resolution imaging over a small field of view and is suitable for OCT as well as other scanning microscopies. Large field of view imaging for screening or surveillance applications can also be achieved by proximally pulling back or advancing the capsule while scanning the distal high-speed micromotor. Circumferential en face OCT was demonstrated in living swine at 250 Hz frame rate and 1 MHz A-scan rate using a MEMS tunable VCSEL light source at 1300 nm. Cross-sectional and en face OCT views of the upper and lower gastrointestinal tract were generated with precision distal pneumatic longitudinal actuation as well as proximal manual longitudinal actuation. These devices could enable clinical studies either as an adjunct to endoscopy, attached to an endoscope, or as a swallowed tethered capsule for non-endoscopic imaging without sedation. The combination of ultrahigh speed imaging and distal scanning capsule technology could enable both screening and surveillance applications.

Depth-encoded all-fiber swept source polarization sensitive OCT.

Polarization sensitive optical coherence tomography (PS-OCT) is a functional extension of conventional OCT and can assess depth-resolved tissue birefringence in addition to intensity. Most existing PS-OCT systems are relatively complex and their clinical translation remains difficult. We present a simple and robust all-fiber PS-OCT system based on swept source technology and polarization depth-encoding. Polarization multiplexing was achieved using a polarization maintaining fiber. Polarization sensitive signals were detected using fiber based polarization beam splitters and polarization controllers were used to remove the polarization ambiguity. A simplified post-processing algorithm was proposed for speckle noise reduction relaxing the demand for phase stability. We demonstrated systems design for both ophthalmic and catheter-based PS-OCT. For ophthalmic imaging, we used an optical clock frequency doubling method to extend the imaging range of a commercially available short cavity light source to improve polarization depth-encoding. For catheter based imaging, we demonstrated 200 kHz PS-OCT imaging using a MEMS-tunable vertical cavity surface emitting laser (VCSEL) and a high speed micromotor imaging catheter. The system was demonstrated in human retina, finger and lip imaging, as well as ex vivo swine esophagus and cardiovascular imaging. The all-fiber PS-OCT is easier to implement and maintain compared to previous PS-OCT systems and can be more easily translated to clinical applications due to its robust design.

Assessment of breast pathologies using nonlinear microscopy.

Rapid intraoperative assessment of breast excision specimens is clinically important because up to 40% of patients undergoing breast-conserving cancer surgery require reexcision for positive or close margins. We demonstrate nonlinear microscopy (NLM) for the assessment of benign and malignant breast pathologies in fresh surgical specimens. A total of 179 specimens from 50 patients was imaged with NLM using rapid extrinsic nuclear staining with acridine orange and intrinsic second harmonic contrast generation from collagen. Imaging was performed on fresh, intact specimens without the need for fixation, embedding, and sectioning required for conventional histopathology. A visualization method to aid pathological interpretation is presented that maps NLM contrast from two-photon fluorescence and second harmonic signals to features closely resembling histopathology using hematoxylin and eosin staining. Mosaicking is used to overcome trade-offs between resolution and field of view, enabling imaging of subcellular features over square-centimeter specimens. After NLM examination, specimens were processed for standard paraffin-embedded histology using a protocol that coregistered histological sections to NLM images for paired assessment. Blinded NLM reading by three pathologists achieved 95.4% sensitivity and 93.3% specificity, compared with paraffin-embedded histology, for identifying invasive cancer and ductal carcinoma in situ versus benign breast tissue. Interobserver agreement was κ = 0.88 for NLM and κ = 0.89 for histology. These results show that NLM achieves high diagnostic accuracy, can be rapidly performed on unfixed specimens, and is a promising method for intraoperative margin assessment.

Ultrahigh speed endoscopic optical coherence tomography for gastroenterology.

We describe an ultrahigh speed endoscopic swept source optical coherence tomography (OCT) system for clinical gastroenterology using a vertical-cavity surface-emitting laser (VCSEL) and micromotor imaging catheter. The system had a 600 kHz axial scan rate and 8 µm axial resolution in tissue. Imaging was performed with a 3.2 mm diameter imaging catheter at 400 frames per second with a 12 µm spot size. Three-dimensional OCT (3D-OCT) imaging was performed in patients with a cross section of pathologies undergoing upper and lower endoscopy. The use of distally actuated imaging catheters enabled OCT imaging with more flexibility, such as volumetric imaging in the small intestine and the assessment of hiatal hernia using retroflex imaging. The high rotational scanning stability of the micromotor enabled 3D volumetric imaging with micron scale volumetric accuracy for both en face OCT and cross-sectional imaging, as well as OCT angiography (OCTA) for 3D visualization of subsurface microvasculature. The ability to perform both structural and functional 3D OCT imaging in the GI tract with microscopic accuracy should enable a wide range of studies and enhance the sensitivity and specificity of OCT for detecting pathology.

Choriocapillaris and choroidal microvasculature imaging with ultrahigh speed OCT angiography.

We demonstrate in vivo choriocapillaris and choroidal microvasculature imaging in normal human subjects using optical coherence tomography (OCT). An ultrahigh speed swept source OCT prototype at 1060 nm wavelengths with a 400 kHz A-scan rate is developed for three-dimensional ultrahigh speed imaging of the posterior eye. OCT angiography is used to image three-dimensional vascular structure without the need for exogenous fluorophores by detecting erythrocyte motion contrast between OCT intensity cross-sectional images acquired rapidly and repeatedly from the same location on the retina. En face OCT angiograms of the choriocapillaris and choroidal vasculature are visualized by acquiring cross-sectional OCT angiograms volumetrically via raster scanning and segmenting the three-dimensional angiographic data at multiple depths below the retinal pigment epithelium (RPE). Fine microvasculature of the choriocapillaris, as well as tightly packed networks of feeding arterioles and draining venules, can be visualized at different en face depths. Panoramic ultra-wide field stitched OCT angiograms of the choriocapillaris spanning ∼32 mm on the retina show distinct vascular structures at different fundus locations. Isolated smaller fields at the central fovea and ∼6 mm nasal to the fovea at the depths of the choriocapillaris and Sattler's layer show vasculature structures consistent with established architectural morphology from histological and electron micrograph corrosion casting studies. Choriocapillaris imaging was performed in eight healthy volunteers with OCT angiograms successfully acquired from all subjects. These results demonstrate the feasibility of ultrahigh speed OCT for in vivo dye-free choriocapillaris and choroidal vasculature imaging, in addition to conventional structural imaging.

Swept source optical coherence microscopy using a 1310 nm VCSEL light source.

We demonstrate high speed, swept source optical coherence microscopy (OCM) using a MEMS tunable vertical cavity surface-emitting laser (VCSEL) light source. The light source had a sweep rate of 280 kHz, providing a bidirectional axial scan rate of 560 kHz. The sweep bandwidth was 117 nm centered at 1310 nm, corresponding to an axial resolution of 13.1 µm in air, corresponding to 8.1 µm (9.6 µm spectrally shaped) in tissue. Dispersion mismatch from different objectives was compensated numerically, enabling magnification and field of view to be easily changed. OCM images were acquired with transverse resolutions between 0.86 µm - 3.42 µm using interchangeable 40X, 20X and 10X objectives with ~600 µm x 600 µm, ~1 mm x 1 mm and ~2 mm x 2 mm field-of-view (FOV), respectively. Parasitic variations in path length with beam scanning were corrected numerically. These features enable swept source OCM to be integrated with a wide range of existing scanning microscopes. Large FOV mosaics were generated by serially acquiring adjacent overlapping microscopic fields and combining them in post-processing. Fresh human colon, thyroid and kidney specimens were imaged ex vivo and compared to matching histology sections, demonstrating the ability of OCM to image tissue specimens.