Cutaneous neurofibromas (cNF) appear commonly in neurofibromatosis type I (NF1) patients. cNFs usually appear in the skin at puberty and proliferate with increasing numbers and sizes throughout life. If cNFs can be detected and treated in their nascent stage, patient’s quality of life can be improved. To detect early-stage cNF, we have employed spatial frequency domain imaging (SFDI) and optical coherence tomography (OCT). SFDI has been applied to screen large skin areas on eleven cNF subjects. Suspect lesions invisible to the unaided eye but detected using SFDI were imaged with OCT to observe lesion microstructure. Three lesions were biopsied to compare with SFDI and OCT images. Suspect nascent cNFs which are invisible to the unaided eye were detected as low optical scattering regions in all patients. Large area screening using SFDI confirmed scattering contrast between the suspect nascent cNF and the surrounding uninvolved skin. Abnormal disc-shaped structures with reduced scattering regions detected by SFDI were also observed in OCT cross-sections.
Coronary chronic total occlusions (CTOs) are severe blockages formed by lipid, fibrous, and calcific material within the artery, halting blood flow for at least three months. Treating CTOs using true-lumen crossing is challenging due to their composition and high tortuosity in coronary arteries. Our study uses intracardiac echocardiography (ICE) catheters to image coronary arteries, introducing a novel 2D and 3D outlining technique. This advancement may improve percutaneous coronary interventions (PCI) for CTOs by providing live imaging feedback during true-lumen crossing procedures, enhancing treatment outcomes.
Recent studies suggest that cavitation effect following laser induced vapor bubble collapse is more dominant than the photothermal effect in stone ablation during laser lithotripsy. Our research aims to introduce an experimental study design that precisely measures each effect's contribution using gypsum phantom stones. To isolate the cavitation-only mechanism after the collapse of the laser-induced vapor bubble, a phantom stone was submerged in a dye solution. The dye solution absorbed all laser light, generating cavitation, with additional experiments confirming the absence of any photothermal effect when the dye was not used. The fiber was positioned both parallel to the stone surface and perpendicular at a 1mm distance, exposing it solely to cavitation. In another set of experiments, a phantom stone was submerged in water and 2μm light from a thulium yttrium aluminum garnet (Tm:YAG) laser was delivered via the same optical fiber positioned (this time) perpendicular to the stone surface. In this case, both optical absorption and cavitation effect from laser-induced vapor bubble collapses were observed but the measured pressure transients showed significantly lower peak pressures compared to the first set. In a final set of experiments, these conditions remained constant, except the fiber was positioned parallel to the stone surface, once again exposing it to only the cavitation from the collapse of the laser induced vapor bubble. Craters created by all methods were imaged using an optical coherence tomography (OCT) system. Measured volumes showed that stone ablation was dominated by photo-thermal, and not by cavitation from the vapor bubble collapse. In fact, in two of the three trials of stone experiments (n=5, each trial) that were subjected to cavitation-only, there was no observable ablation. One trial produced an average volume that was 50% smaller than the average resulting from a single photo-thermal-only case (p = 0.0022 < 0.05). Our results suggest that finetuning of lithotripsy procedures with focus on energy transmission to the stone can provide optimal results.
Pneumatic control devices were constructed to apply negative pressure on targeted human skin areas to increase or decrease blood flow for improving laser therapy. The effect of pneumatic control on blood flow in a cutaneous skin region was evaluated using optical coherence tomography-angiography (OCT-A). Dynamic changes in blood flow under the application of negative pressure to the dorsal hand and forearm skin were confirmed in acquired OCT-A images. We also observed the device size-dependent difference in blood flow change. Results suggest the feasibility of bi-directional blood flow control using a pneumatic control device.
A major constraint in photomedicine is the scattering of photons within tissue, which can limit the penetration of light to reach target structures such as a tumor. Laser-induced shockwaves could offer a potential approach to mitigate this constraint by modifying the optical scattering properties of tissue and re-directing the light to targeted regions. Preliminary results in a skin phantom suggest that laser-induced shockwaves can alter optical characteristics of a turbid medium, and subsequently, induce changes in light propagation.
SignificanceTraditional pathology workflow suffers from limitations including biopsy invasiveness, small fraction of large tissue samples being analyzed, and complex and time-consuming processing.AimWe address limitations of conventional pathology workflow through development of a laser microbiopsy device for minimally invasive harvest of sub-microliter tissue volumes. Laser microbiopsy combined with rapid diagnostic methods, such as virtual hematoxylin and eosin (H&E) imaging has potential to provide rapid minimally invasive tissue diagnosis.ApproachLaser microbiopsies were harvested using an annular shaped Ho:YAG laser beam focused onto the tissue surface. As the annulus was ablated, the tissue section in the center of the annulus was ejected and collected directly onto a glass slide for analysis. Cryogen spray cooling was used before and after laser harvest to limit thermal damage. Microbiopsies were collected from porcine skin and kidney. Harvested microbiopsies were imaged with confocal microscopy and digitally false colored to provide virtual H&E images.ResultsMicrobiopsies were successfully harvested from porcine skin and kidney. Computational and experimental results show the benefit of cryogen pre- and post-cooling to limit thermal damage. Virtual H&E images of microbiopsies retained observable cellular features including cell nuclei.ConclusionsLaser microbiopsy with virtual H&E imaging shows promise as a potential rapid and minimally invasive tool for biopsy and diagnosis.
Tissue biopsy and histological evaluation is the gold standard for disease diagnosis including cancer. For example, a punch several millimeters in diameter is often used to biopsy suspicious skin sites. The biopsy is then formalin fixed, paraffin embedded, sectioned, stained with hematoxylin and eosin (H&E), and examined by a pathologist. While this process has been the gold standard for decades, two limitations are recognized. First, the biopsy is invasive with a limited number that can be reasonably tolerated by the patient. Second, the tissue processing steps are slow. We report an alternative approach consisting of a laser microbiopsy for harvest of sub-microliter (<1 mm3) tissue sections combined with rapid virtual H&E staining methods. A Ho:YAG laser (Lumenis P120) was shaped into an annular beam and focused onto ex vivo porcine skin. The epidermis and dermis were laser cut and the tissue section in the center of the annulus was ejected and collected by an overlying glass coverslip. Tetrafluoroethane (R134A) was sprayed at the ablation site prior to ablation and at the collected tissue section post ablation to limit thermal damage and preserve histological features. Two virtual H&E imaging methods were tested with confocal microscopy. The first combined acridine orange fluoresce with reflectance. The second combined acridine orange and sulforhodamine 101 fluorescence. For each method, the two channels were false colored and combined to create virtual H&E images. Virtual H&E images show histological features, including cell nuclei. Laser microbiopsy is minimally invasive, harvesting tissue sections on the order of 0.01 to 0.1 mm3, and tissue processing is rapid requiring 2 min or less for staining. Laser microbiopsy is a promising candidate technique for rapid minimally invasive diagnosis.
Coronary chronic total occlusions (CTOs) are atherosclerotic plaques comprised of lipid, fibrous and hard calcific material that originate in the vessel wall and extend into the lumen, restricting luminal cross-section by 100% resulting in complete stoppage of blood flow in the affected artery for at least three months. Due to their structure and calcific composition, CTOs are very difficult to treat with existing percutaneous coronary interventional (PCI) techniques. CTOs frequently have a hard fibro-calcific cap on the proximal side with a softer lipidic composition in the interior and distal side. We constructed a novel catheter system with a fiber coupled Ho:YAG laser (2.1um, Coherent Inc) for cutting and a biocompatible CO2 cooling system delivered through a 200um conduit for intravascular cooling. Laser radiation delivered a maximum average power of 20W corresponding to a pulse energy of 300mJ, pulse duration of 200μs, and a pulse repetition rate of 10Hz. Light emitted from the fiber was directed onto ex vivo suspect-calcified CTO arteries (n=3). Successful CTO crossing was achieved in all ex vivo samples. Histological processing showed greater than 50% reduction in residual thermal damage in crossed CTO regions with CO2 cooling compared to no cooling. The miniature device was also used to cross CTOs in an in vivo rabbit femoral CTO model (n=4) under Xray fluoroscopy guidance and subsequent contrast angiography confirmed restoration of blood flow.
Percutaneous coronary intervention (PCI) for improving calcified coronary artery compliance remains a challenge and is associated with high rates of complications and adverse outcomes. In addition to traditional rotational atherectomy devices for improving coronary artery compliance, recently, electric discharge plasma mediated shockwave therapy has been introduced to cause calcium fracture and improve coronary compliance. However, this intervention has cardiac pacing limitations. Laser lithotripsy is commonly utilized to fracture kidney stones. High powered laser pulses are transmitted via small diameter optical fibers (200-400 μm core diameter) to the stone surface, where they induce fracture. We implemented a novel catheter device that utilizes indocyanine green (ICG) filled balloon to produce calcium fractures. At 2mg/mL, ICG has greater than 5x higher absorption coefficient (256cm-1, 755nm) than water at 2.1 μm, a typical target of holmium lasers (~40cm-1, 2.1μm) during lithotripsy. To demonstrate the feasibility of laser induced calcium fracture a balloon catheter device (2mm outer diameter un-inflated, 1 meter long) was constructed with a fiber port coupled to alexandrite lasers (755nm) and a balloon port to fill biocompatible ICG in front of the fiber. Different temporal pulse regimes (millisecond to sub-nanosecond) were explored inducing shockwaves pressure amplitudes higher than 50 atm sufficient to cause fracture in coronary artery phantoms made from Ultracal 30® material. This approach does not require cardiac pacing and can markedly improve arterial vessel compliance during stent deployment.
Lasers are commonly employed in surgery for hard and soft tissues due to their precise space-time energy delivery and compatibility with optical fibers for delivery into body cavities, including for treatment of urological diseases. Infrared laser ablation in tissues can result in non-specific heating and thermal injury. Methods that maximize ablation efficiency, or tissue volume removed per unit energy, while minimizing non-specific thermal injury can improve surgical workflows and outcomes. We report a novel approach for increased ablation efficiency by modifying the beam shape. Specifically, a Ho:YAG laser is shaped into a converging annular beam. Ablation efficiency was measured on a hard tissue phantom (BegoStone) and soft tissue (porcine kidney). An annular beam ~800 μm in diameter was used to ablate each sample at 10 different locations using a single 1 J pulse per location. The procedure was repeated using a circular beam with similar diameter by placing a 200 μm fiber 1 mm from the tissue surface. Each ablation crater was imaged with optical coherence tomography and the crater volumes calculated from recorded images. For hard tissue phantoms, ablation efficiency increased 183% for annular vs. circular beams (0.065±0.013 vs. 0.023 ± 0.003 mm3 /J). For soft tissue, ablation efficiency increased 69% for annular vs. circular beams (0.098±0.021 vs 0.058 ± 0.018 mm3 /J). Hard and soft tissue ablation with an annular beam is a promising technique for increasing the speed and safety of laser surgery.
Laser generated bubbles surrounding plasmonic nanoparticles (known as plasmonic nanobubbles) are an attractive candidate approach for imaging and therapy, especially for cancer research. Nanobubbles are typically small in size (in microns), have a short lifetime (microseconds), and require complex instrumentation to characterize. Current approaches measure the bubble size using flash photography or an ultrafast camera and the bubble lifetime indirectly with a light scattering of a probe beam. Here, we demonstrate swept-source (1310+/-70 nm, 100 Khz scan rate) optical coherence tomography (OCT) as a new approach to monitoring plasmonic nanobubbles' dynamics. We recorded a one-dimensional (A-scan) analysis on a thin Au-nanoparticle-embedded gelatin layer and monitored the phase offset of the gelatin/air interface where nanobubbles were generated in response to a high energy single 10-ns 1064nm Nd: YAG laser pulse. We observed the gel/air interface movement reflecting the formation and collapse of nanobubbles with radius in the range of 200 – 600 nm and lifetime up to 100 microseconds corresponding to an incident laser fluence of 0.9 – 2.4J/cm2. We observed interface oscillations following nanobubble collapse. In conclusion, OCT may provide a simple technique to characterize both nanobubble's size and lifetime in response to pulsed laser irradiation of plasmonic nanoparticles.
Tissue sampling is required for disease diagnostics and research. Traditional tissue sampling tools often remove more tissue than required, causing unnecessary pain and morbidity. We report a minimally invasive laser microbiopsy system to sample sub-microliter (<1mm3 ) tissue sections. A focused annular beam cuts conical-shaped tissue sections. Compatible diagnostic methods may include standard histopathology, rapid point-of-care microscopy, and genetic testing. An objective of the approach is to harvest a minimal volume while limiting damage to retain tissue integrity for diagnosis. The laser microbiopsy system consists of a Ho:YAG laser (Lumenis P120), a ZnSe aspheric collimator (ISP optics, f = 50.8 mm), two fused silica axicons (Thorlabs, α = 20°), and an aspheric focusing lens (ISP optics, f = 25.4 mm). The system is characterized to evaluate the required laser dosimetry, range of harvested tissue volumes, and residual thermal damage. We first characterized the system in silico using raytracing (Zemax) and a heat transfer modeling. We then characterized the system experimentally performing micro-biopsies in ex vivo porcine skin. Raytracing and experimental measurements show agreement in annular beam shape with an inner and outer radius of 300 μm and 400 μm near the beam focus. Experimental results show ability to harvest tissue sections ranging from 0.009 to 0.10 mm3 using pulse energies from 1 - 2 J.
Optical coherence tomography (OCT) has shown promise as a process sensor in selective laser sintering (SLS) due to its ability to yield depth-resolved data not attainable with conventional sensors. However, OCT images of nylon 12 powder and nylon 12 components fabricated via SLS contain artifacts that have not been previously investigated in the literature. A better understanding of light interactions with SLS powder and components is foundational for further research expanding the utility of OCT imaging in SLS and other additive manufacturing (AM) sensing applications. Specifically, in this work, nylon powder and sintered parts were imaged in air and in an index matching liquid. Subsequent image analysis revealed the cause of “signal-tail” OCT image artifacts to be a combination of both inter and intraparticle multiple-scattering and reflections. Then, the OCT imaging depth of nylon 12 powder and the contrast-to-noise ratio of a sintered part were improved through the use of an index matching liquid. Finally, polymer crystals were identified as the main source of intraparticle scattering in nylon 12 powder. Implications of these results on future research utilizing OCT in SLS are also given.
Neurological cancer surgeries require specialized tools that enhance imaging for precise cutting and removal of tissue without damaging adjacent neurological structures. The novel combination of high-resolution fast optical coherence tomography (OCT) alongside short pulsed nanosecond thulium (Tm) lasers offers stark advantages utilizing the superior beam quality, high volumetric tissue removal rates of thulium lasers with minimal residual thermal footprint in the tissue and avoiding damage to delicate sub-surface structures (e.g., nerves and microvessels); which has not been showcased before. A bench-top system is constructed, using a 15W 1940nm nanosecond pulsed Tm fiber laser (500uJ pulse energy, 100ns pulse duration, 30kHz repetition rate) for removing tissue and a swept source laser (1310±70nm, 100kHz sweep rate) is utilized for OCT imaging, forming a combined Tm/OCT system – a smart laser knife. The OCT image-guidance informs the Tm laser for cutting/removal of targeted tissue structures. Tissue phantoms were constructed to demonstrate surgical incision with blood vessel avoidance on the surface where 2mm wide 600um deep cuts are executed around the vessel using OCT to guide the procedure. Cutting up to delicate subsurface blood vessels (2mm deep) is demonstrated while avoiding damage to their walls. A tissue removal rate of 5mm^3/sec is obtained from the bench-top system. We constructed a blow-off model to characterize Tm cut depths taking into account the absorption coefficients and beam delivery systems to compute Arrhenius damage integrals. The model is used to compare predicted tissue removal rate and residual thermal injury with experimental values in response to Tm laser-tissue modification.
State-of-the-art treatment for joint diseases like osteoarthritis focus on articular cartilage repair/regeneration by stem cell implantation therapy. However, the technique is limited by a lack of precision in the physician’s imaging and cell deposition toolkit. We describe a novel combination of high-resolution, rapid scan-rate optical coherence tomography (OCT) alongside a short-pulsed nanosecond thulium (Tm) laser for precise cell seeding in cartilage. The superior beam quality of thulium lasers and wavelength of operation 1940 nm offers high volumetric tissue removal rates and minimizes the residual thermal footprint. OCT imaging enables targeted micro-well placement, precise cell deposition, and feature contrast. A bench-top system is constructed using a 15 W, 1940 nm, nanosecond-pulsed Tm fiber laser (500 μJ pulse energy, 100 ns pulse duration, 30kHz repetition rate) for removing tissue, and a swept source laser (1310 ± 70 nm, 100 kHz sweep rate) for OCT imaging, forming a combined Tm/OCT system – a “smart laser knife”. OCT assists the smart laser knife user in characterizing cartilage to inform micro-well placement. The Tm laser creates micro-wells (2.35 mm diameter length, 1.5 mm width, 300 μm deep) and micro-incisions (1 mm wide, 200 μm deep) while OCT image-guidance assists and demonstrates this precision cutting and cell deposition with real-time feedback. To test micro-well creation and cell deposition protocol, gelatin phantoms are constructed mimicking cartilage optical properties and physiological structure. Cell viability is then assessed to illustrate the efficacy of the hydrogel deposition. Automated OCT feedback is demonstrated for cutting procedures to avoid important surface/subsurface structures. This bench-top smart laser knife system described here offers a new image-guided approach to precise stem cell seeding that can enhance the efficacy of articular cartilage repair.
We present development of a nanosecond Q-switched Tm3+-doped fiber laser with 16 W average power and 4.4 kW peak power operating at 1940 nm. The laser has a master oscillator power amplifier design, and uses large mode area Tm3+-doped fibers as the gain medium. Special techniques are used to splice Tm3+-doped fibers to minimize splice loss. The laser design is optimized to reduce non-linear effects, including modulation instability. Pulse width broadening due to high gain is observed and studied in detail. Medical surgery is a field of application where this laser may be able to improve clinical practice. The laser together with scanning galvanometer mirrors is used to cut precisely around small footprint vessels in tissue phantoms without leaving any visible residual thermal damage. These experiments provide proof-of-principle that this laser has promising potential in the laser surgery application space.
Optical coherence tomography (OCT) retinal imaging contributes to understanding central nervous system (CNS)
diseases because the eye is an anatomical “window to the brain” with direct optical access to nonmylenated retinal
ganglion cells. However, many CNS diseases are associated with neuronal changes beyond the resolution of standard
OCT retinal imaging systems. Though studies have shown the utility of scattering angle resolved (SAR) OCT for particle
sizing and detecting disease states ex vivo, a compact SAR-OCT system for in vivo rodent retinal imaging has not
previously been reported. We report a fiber-based SAR-OCT system (swept source at 1310 nm ± 65 nm, 100 kHz scan
rate) for mouse retinal imaging with a partial glass window (center aperture) for angular discrimination of backscattered
light. This design incorporates a dual-axis MEMS mirror conjugate to the ocular pupil plane and a high collection
efficiency objective. A muring retina is imaged during euthanasia, and the proposed SAR-index is examined versus time.
Results show a positive correlation between the SAR-index and the sub-cellular hypoxic response of neurons to
isoflurane overdose during euthanasia. The proposed SAR-OCT design and image process technique offer a contrast
mechanism able to detect sub-resolution neuronal changes for murine retinal imaging.
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