This PDF file contains the front matter associated with SPIE Proceedings Volume 8565, including the Title Page, Copyright information, Table of Contents, and the Conference Committee listing.

Laser ablation may be a promising method for removal of skin lesions, with the potential for better cosmetic outcomes and reduced scarring and infection. An obstacle to implementing laser ablation is that the treatment leaves no tissue for histopathological analysis. Pre-operative and intra-operative mapping of BCCs using confocal microscopy may guide the ablation of the tumor until all tumor is removed. We demonstrate preliminary feasibility of confocal microscopy to guide laser ablation of BCCs in freshly excised tissue from Mohs surgery. A 2940 nm Er:YAG laser provides efficient ablation of tumor with reduced thermal damage to the surrounding tissue.

Reflectance Confocal Microscopic (RCM) imaging of obliquely-oriented optical sections, rather than with traditional z-stacks, shows depth information that more closely mimics the appearance of skin in orthogonal sections of histology. This approach may considerably reduce the amount of data that must be acquired and processed. However, as with z-stacks, purely visual detection of the dermal-epidermal junction (DEJ) in oblique images remains challenging. Here, we have extended our original algorithm for localization of DEJ in z-stacks to oblique images. In addition, we developed an algorithm for detecting wrinkles, which in addition to its intrinsic merit, gives useful information for DEJ detection.

The effects on skin of two commercially available topical creams for the treatment of eczema are quantitatively studied using optical coherence tomography. An archetypal corticosteroid (Betamethasone valerate) is compared with a nonsteroidal anti-inflammatory drug (Tacrolimus monohydrate) via left/right comparisons of the epidermal thickness of volar forearm skin on selected volunteers, at baseline and after 14 days of treatment. In 3 of 4 subjects we confirmed previous observations that corticosteroids produce pronounced physical thinning of the epidermis over timescales of a few weeks. In 3 of 4 subjects we further found that Tacrolimus produced no change in epidermal thickness. In one of 4 subjects we found evidence that the epidermis was actually thickened following treatment using Tacrolimus.

A compact and easy-to-handle photocoagulation device was used for inducing an immediate coagulation effect in skin large superficial abrasions, reducing the recovering time and improving the wound healing process. The handheld illumination device consists of a high power LED, emitting in the blue region of the spectrum, mounted in a suitable and ergonomic case, together with power supply, electronics, and batteries. The working principle of the LED-based photocoagulator is a photothermal effect: the blue light is selectively absorbed by the haemoglobin content of the blood and then converted into heat. Here we present an in vivo study performed on animal models. 10 Sprague Dawley rats (Harlan, Italy, weighing 200-250 g) were used to study the wound healing process. On the back of each rat, four large abrasions were mechanically produced: two of them were used as a control, while the other two were treated with the photocoagulator, keeping it at a constant distance (2 mm) from the target, in continuous slow motion (treatment time: tens of seconds). The induced photothermal effect was monitored by an infrared thermocamera in order to avoid accidental thermal damage and to control the temperature dynamics during treatment. Objective observations, histopathological analysis and non-linear microscopy performed in a 8 days follow-up study showed no adverse reactions and no thermal damage in the treated areas and surrounding tissues. Moreover, a faster healing process and a better recovered morphology was evidenced in the treated tissue.

Diffuse reflectance spectra (DRS) of biological samples are commonly measured using an integrating sphere (IS), in which spectrally broad illumination light is multiply scattered and homogenized. The measurement begins by placing a highly reflective white standard against the IS sample opening and collecting the reflected light at the signal output port to account for illumination field. After replacing the white standard with test sample of interest, DRS of the latter is determined as the ratio of the two values at each involved wavelength. However, because test samples are invariably less reflective than the white standard, such a substitution modifies the illumination field inside the IS. This leads to underestimation of the sample’s reflectivity and distortion of measured DRS, which is known as single-beam substitution error (SBSE). Barring the use of much more complex dual-beam experimental setups, involving dedicated IS, literature states that only approximate corrections of SBSE are possible, e.g., by using look-up tables generated with calibrated low-reflectivity standards. We present a practical way to eliminate the SBSE using IS equipped with an additional “reference” output port. Two additional measurements performed at this port (of the white standard and sample, respectively) namely enable an accurate compensation for above described alteration of the illumination field. In addition, we analyze the dependency of SBSE on sample reflectivity and illustrate its impact on measurements of DRS in human skin with a typical IS.

Collagen represents major protein component of the extracellular matrix (ECM), thus its monitoring could be used as an ideal tool for assessing wound healing following therapy. Presently, there is a great need to develop quick, objective, non-destructive method to monitor collagen synthesis during progression of wound healing. The applicability of the Laser induced fluorescence technique towards wound healing monitoring by measuring collagen levels at different stages of the healing progression is the main idea behind the current work. Six to eight weeks old Swiss albino mice with ccircular wounds of 15 mm diameter were illuminated with single exposure of 2 J/cm² from He-Ne laser (632.8 nm; 7 mW power; 4.02 mW/cm² power density) along with un-illuminated and un-wounded controls. Spectroscopic changes were monitored by recording in vivo fluorescence from each animal (under anesthesia) at different post-wounding days (5^th, 10^th, 30^thday) by exciting granulation tissue/skin with 325 nm He-Cd laser. The autofluorescence from the tissue/skin was recorded from four different sites and four spectra were recorded from each site. A total of 2160 spectra were recorded from 45 animals. The in vivo fluorescence studies have shown significant increase (P<0.001) in collagen synthesis upon treatment with optimum laser dose of 2 J/cm² immediately after wounding as compared to un-illuminated control group. It can conclude that in vivo fluorescence measurement is effective in monitoring wound healing and hence could be used over ex vivo method as an objective and non-intrusive method to monitor collagen levels

The healing process of chronic wounds is complex, and the complete pathogenesis is not known. Diagnosis is currently based on visual inspection, biopsies and collection of samples from the wound surface. This is often time consuming, expensive and to some extent subjective procedures. Hyperspectral imaging has been shown to be a promising modality for optical diagnostics. The main objective of this study was to identify a suitable technique for reproducible classification of hyperspectral data from a wound and the surrounding tissue. Two statistical classification methods have been tested and compared to the performance of a dermatologist. Hyperspectral images (400-1000 nm) were collected from patients with venous leg ulcers using a pushbroom-scanning camera (VNIR 1600, Norsk Elektro Optikk AS).Wounds were examined regularly over 4 - 6 weeks. The patients were evaluated by a dermatologist at every appointment. One patient has been selected for presentation in this paper (female, age 53 years). The oxygen saturation of the wound area was determined by wavelength ratio metrics. Spectral angle mapping (SAM) and k-means clustering were used for classification. Automatic extraction of endmember spectra was employed to minimize human interaction. A comparison of the methods shows that k-means clustering is the most stable method over time, and shows the best overlap with the dermatologist’s assessment of the wound border. The results are assumed to be affected by the data preprocessing and chosen endmember extraction algorithm. Results indicate that it is possible to develop an automated method for reliable classification of wounds based on hyperspectral data.

Arthritis is a chronic inflammatory disease that induces potentially damaging and commonly disabling. Various imaging modalities have been used for the evaluation of arthritis. This study aimed to investigate the feasibility of laser speckle contrast image (LSCI) in the evaluation of the severity and early stage of arthritis in animal model. Arthritis was induced on mouse foot and evaluated by a trained expert and the LSCI. The arthritis severity was quantitatively evaluated by speckle index (SI) computed from LSCI. In visual inspection by an expert, it was difficult to evaluate the arthritis because there was no noticeable different between control mouse group (CMG) and arthritis mouse group (AMG) in erythema. However, arthritis was easily evaluated by significant SI different between the CMG and AMG. In addition, the LSCI also successfully evaluated the early stage of arthritis, presenting different SI distribution depending on lesion.

Background and Objective: Spatial Frequency Domain Imaging (SFDI) is a non-contact wide-field optical imaging technology currently being developed to investigate the feasibility of quantitative non-invasive evaluation of burn wound severity in a rat model. Our objective is to determine the potential of SFDI for mapping quantitative changes in spatially resolved tissue oxygen saturation and water concentration may be indicative of burn wound severity, healing, and further complications. In this portion of the investigation, we focus on the development of a rat burn model and the acute response of tissue to burn wounds. Study Design/Materials and Methods: A controlled burn protocol involving a heated brass comb was applied to 6 rats. Imaging was acquired at 17 evenly spaced wavelengths in the near-infrared from 650 to 970 nm. Over the course of the 3 hour post-burn period, we were able to map quantitative changes in spatially resolved chromophores. Burn severities were verified post-experiment using standard H and E histology and optical microscopy. Results/Conclusion: In total, we were able to induce 12 superficial-partial thickness burns, 8 deep-partial thickness burns, and 4 full thickness burns in our rat models. While several tissue chromophores were tracked, we found that changes in oxygen saturation and water concentration to be sensitive indicators of burn severity. Future work will include additional longitudinal studies over a period of days in order to investigate which parameters are correlated to tissue healing.

Normal skin barrier function depends on having a viable epidermis, an epithelial layer formed by keratinocytes. The transparent epidermis, which is less than a 100 mum thick, is nearly impossible to see. Thus, the clinical evaluation of re-epithelialization is difficult, which hinders selecting appropriate therapy for promoting wound healing. An imaging system was developed to evaluate epithelialization by detecting endogenous fluorescence emissions of cellular proliferation over a wide field of view. A custom-made 295 nm ultraviolet (UV) light source was used for excitation. Detection was done by integrating a near-UV camera with sensitivity down to 300 nm, a 12 mm quartz lens with iris and focus lock for the UV regime, and a fluorescence bandpass filter with 340 nm center wavelength. To demonstrate that changes in fluorescence are related to cellular processes, the epithelialization of a skin substitute was monitored in vitro. The skin substitute or construct was made by embedding microscopic live human skin tissue columns, 1 mm in diameter and spaced 1 mm apart, in acellular porcine dermis. Fluorescence emissions clearly delineate the extent of lateral surface migration of keratinocytes and the total surface covered by the new epithelium. The fluorescence image of new epidermis spatially correlates with the corresponding color image. A simple, user-friendly way of imaging the presence of skin epithelium would improve wound care in civilian burns, ulcers and surgeries.

Scarring can lead to significant cosmetic, psychosocial, and functional consequences in patients with hypertrophic scars from burn and trauma injuries. Therefore, quantitative assessment of scar is needed in clinical diagnosis and treatment. The Vancouver Scar Scale (VSS), the accepted clinical scar assessment tool, was introduced in the nineties and relies only on the physician subjective evaluation of skin pliability, height, vascularity, and pigmentation. To date, no entirely objective method has been available for scar assessment. So, there is a continued need for better techniques to monitor patients with scars. We introduce a new spectral imaging system combining out-of-plane Stokes polarimetry, Spatial Frequency Domain Imaging (SFDI), and three-dimensional (3D) reconstruction. The main idea behind this system is to estimate hemoglobin and melanin contents of scar using SFDI technique, roughness and directional anisotropy features with Stokes polarimetry, and height and general shape with 3D reconstruction. Our proposed tool has several advantages compared to current methodologies. First and foremost, it is non-contact and non-invasive and thus can be used at any stage in wound healing without causing harm to the patient. Secondarily, the height, pigmentation, and hemoglobin assessments are co-registered and are based on imaging and not point measurement, allowing for more meaningful interpretation of the data. Finally, the algorithms used in the data analysis are physics based which will be very beneficial in the standardization of the technique. A swine model has also been developed for hypertrophic scarring and an ongoing pre-clinical evaluation of the technique is being conducted.

A few new low invasive fiber laser technologies for treatment of 1) capillary malformations (port-wine stains), 2) venous, arterial, and arteriovenous malformations, 3) lymphatic malformations of 3 types: micro, small and large-cystic lymphangiomas are presented in this work. There were applied 1.56 μm laser distant photocoagulation, 1.56 μm laser endovascular thermotherapy, 1.9 μm laser instant ablation, 1.9 μm laser interstitial thermotherapy, and 1.9 μm laser excision. The technologies were applied to about 300 patients. Good clinical and esthetic results have been achieved in great majority cases.

We report on “eye-safe” erbium glass laser operating at Short-Wave Infra-Red (SWIR) region at 1534nm, to treat Onychomycosis or toenail fungus. Infected toenails of 12 patients were treated over a 3 month period using both long pulse and Q-switched laser output pulses. Our results compared favorably to Neodymium Yittrium Aluminum Garnet (Nd:YAG) laser fungus treatment studies as reported in literature. Nd:YAG laser devices, operating in the Near Infra- Red, (NIR) region at 1064nm, have recently become an effective alternative treatment to traditional oral medications used to treat nail fungal infections. Conventional nail infection treatments employ medications such as allylamines, azoles and other classes of antifungal drugs that are unpopular due to numerous side-affects and drug interactions. Side effects of these drugs include headache, itching, loss of sense of taste, nausea, diarrhea, heart failure and even potential death from liver failure [1,2,3]. The effectiveness of conventional oral antifungal medications varies. In addition, antifungal prescription drugs are administered for long periods ranging from 6 weeks to 18 months. Nd:YAG antifungal laser treatment reports claim high success rates (65-95%) in eradicating toenail fungus and without adverse side-affects. Multiple laser treatments are administered over a 3 to 6 month period [4,5,6,7]. Our initial treatments performed with the Er:glass laser on toenail fungus patients required only 1 to 2 treatments for cure. This same SWIR laser was used in experiments to treat Athlete's Foot fungal infections. The 1534nm Er:glass laser emission has been found to be well optimized for dermatological treatments due high transmission properties of human skin in the SWIR region. Increased depth of tissue penetration is well-tolerated and provides for effective treatment of various skin conditions. [8,9,10,11] “Eye-safe” Class I lasers provide for practical skin and nail tissue treatment without the need for eye-protection goggles. Laser safety filters may inhibit a practitioner’s vision and ability to distinguish skin and nail regions exhibiting different colors and textures. The laser is “eye-safe” due to the fact that Megawatt peak power Q-switched lasers operating at 1.54um in the narrow spectral window between 1.4um and 1.6um are approximately 8000 times more eye-safe than other laser devices operating in the visible and near infrared. Long-pulse or free running lasers operating in this wavelength range are ~ 2000 times more eye-safe [12].

In this paper, we studied the phenylalanine amino acid in different systems, it means, the amino acid with different solvent molecules as water, ammonia and urea, via density functional theory (DFT). These molecules are present in the natural moisturizing factor (NMF) of skin that is found in the stratum corneum. The aim of this study is improving the understanding of the environment role and its influences. Therefore, in this paper we analyzed the vibrational modes (and compared to FT-Raman and FT-IR experimental values) and electronic properties as the charge densities (HOMO and LUMO). We also verified the hydrophobic effect and the dependence of amino acid structure on the quantities of solvent.

This work aimed the development of a discriminating model, using Raman spectroscopy, based on the estimated concentration of biochemical components presented in skin, for in vivo diagnosis. Raman spectra were collected in patients who underwent excision surgery of suspicious lesions at the lesion site and at a normal circumjacent site. It has been estimated the relative amount of selected biochemical compounds presented in skin. The Raman spectra of normal and malignant (basal cell carcinoma - BCC and squamous cell carcinoma - SCC) skin are quite similar, with some spectral differences in the regions of lipids, nucleic acids, and hemoglobin. Some biochemicals showed statistically significant differences among N, BCC and SCC, such as elastin, ceramide, melanin, nucleid acid, actin and phenylalanine. Elastin and ceramide presented significant differences between N and BCC, melanin, DNA and actin presented significant differences between N and BCC and between N and SCC, being melanin and DNA decreased in neoplasias, in contrast with actin, that increased in neoplasias. Concentration of phenylalanine was significantly increased for SCC compared to N and BCC. The relative concentration of melanin, DNA and phenylalanine showed sensitivity, specificity and accuracy of about 81%, 65% and 60%, respectively, using Mahalanobis distance as a discriminator. This model is being incorporated to a Raman system with automated data collection and processing that could be used for a future in vivo, real time discrimination algorithm.

Background: Painful bladder syndrome/interstitial cystitis (PBS/IC) is defined as a syndrome of urgency, frequency, and suprapubic pain in the absence of positive urine culture or obvious bladder pathology. As no specific etiology has been identified yet, no specific methodology exists for diagnosis of this condition. One potential etiology of PBS/IC is inflammation of the bladder mucosa associated with abnormal angiogenesis and ulcerative lesions. The purpose of this study was to examine the feasibility of using transcutaneous near infrared spectroscopy (NIRS) of the bladder to monitor tissue oxygenation and hemodynamics as a means of differentiating subjects diagnosed with PBS/IC from those with other bladder conditions. Methods: Twenty-four adult patients with lower urinary tract dysfunction were divided into 2 groups, PBS/IC and non-PBS/IC after standard diagnostic investigations. Detrusor oxygen saturation percentage (TSI%) was measured in all subjects while they were at rest in a supine position, using a spatially resolved (SR) NIRS instrument. Mean values of detrusor TSI% were significantly different between the two groups (74.2%±4.9 in PBS/IC vs. 63.6%±5.5 in non-PBS/IC, P<0.0005). Results: Noninvasive NIRS interrogation of the bladder demonstrated that patients diagnosed as having PBS/IC had significantly higher detrusor oxygen saturation at rest. Conclusions: SR-NIRS as a feasible non-noninvasive entity for use in the evaluation of patients for the presence or absence of physiologic changes associated with PBS/IC.

The treatment of choice for patients with end-stage renal disease is kidney transplantation. However, acute tubular necrosis (ATN) induced by an ischemic insult (e.g., from prolonged ex vivo storage times, or non-heart beating cadavers) is a major factor limiting the availability of donor kidneys. In addition, ischemic induced ATN is a significant risk factor for eventual graft survival and can be difficult to discern from rejection. Currently, there are no rapid and reliable tests to determine ATN suffered by donor kidneys and whether or not donor kidneys might exhibit delayed graft function. OCT (optical coherence tomography) is a rapidly emerging imaging modality that can function as a type of “optical biopsy”, providing cross-sectional images of tissue morphology in situ and in real-time. In a series of recent clinical trials, we evaluated the ability of OCT to image those features of the renal microstructure that are predictive of ATN. Specifically, we found that OCT could effectively image through the intact human renal capsule and determine the extent of acute tubular necrosis. We also found that Doppler based OCT (i.e., DOCT) revealed renal blood flow dynamics that is also reported to be a determiner of post-transplant renal function. This kind of information will allow transplant surgeons to make the most efficient use of available donor kidneys, eliminate the possible use of bad donor kidneys, provide a measure of expected post-transplant renal function, and allow better distinction between post-transplant immunological rejection and ischemic-induced acute renal failure.

Successful identification of the cavernous nerves (CN’s) during radical prostatectomy requires detection of the CN’s through a thin layer of overlying fascia. This study explores the 1490 nm infrared (IR) diode laser wavelength for rapid and deep subsurface CN stimulation in a rat model, in vivo. A 150-mW, 1490-nm diode laser providing an optical penetration depth of ~ 520 μm was used to stimulate the CN’s in 8 rats through a single mode fiber optic probe with 1-mm-diameter spot and 15 s irradiation time. Successful ONS was judged by an intracavernous pressure response (ICP) in the rat penis. Subsurface ONS at 1490 nm was also compared with previous studies using 1455 and 1550 nm IR diode laser wavelengths. ONS was observed through fascia layers up to 380 μm thick using an incident laser power of ~ 50 mW. ICP response times as short as 4.6 ± 0.2 s were recorded using higher laser powers bust still below the nerve damage threshold. The 1490-nm diode laser represents a compact, low cost, high power, and high quality infrared light source for use in ONS. This wavelength provides deeper optical penetration than 1455 nm and more rapid and efficient nerve stimulation than 1550 nm.

Optical nerve stimulation (ONS) may be useful for intra-operative identification and preservation of the prostate cavernous nerves (CN’s), responsible for erectile function, during radical prostatectomy. ONS relies on a photothermal mechanism of laser-tissue interaction in which elevating nerve temperature to within a narrow range (~ 42 - 47 ^oC) is critical to successful nerve activation without thermal damage. This preliminary study explores a prototype temperature-controlled laser system for maintaining a constant nerve temperature during ONS of the rat prostate CN’s. A 150-mW, 1455-nm diode laser was operated in continuous-wave mode during stimulation of the rat CN’s for 30 s through a fiber optic probe with 1-mm-diameter spot. The all-single-mode ONS system was controlled by a computer which opened and closed an in-line mechanical shutter in response to an IR sensor, with a temperature set-point of 45 ^oC. Thermal camera temperature measurements and ONS without temperature control were performed for comparison. Strong correlation was observed between IR sensor and camera temperatures. With temperature control, CN temperature was maintained at 45.0 ± 1.5 ^oC. Without IR sensor feedback, CN temperatures continued to rise during ONS, reaching unsafe levels of ~ 50 ^oC.

Photo dynamic diagnosis (PDD) is a convenient and well-documented procedure for diagnosis of bladder cancer and tumours using endoscopic techniques. At present, this procedure is available only for routine use in an operating room (OR) and often with substantial photobleaching effects of the photosensitizer. We present a novel optical design of the endoscopic PDD procedure that allows the procedure to be performed in the outpatient department (OPD) and not only in the OR. Thereby, inpatient procedures lasting 1-2 days may be replaced by a few hours lasting procedure in the OPD. Urine blurs the fluorescence during PDD used in the OPD. Urine contains fluorescent metabolites that are excited by blue light giving an opaque green fluorescence confounding the desired red fluorescence (PDD) from the tumour tissue. Measurements from the clinical situation has shown that some systems for PPD based on blue light illumination (PDD mode) and white light illumination used for bladder tumour diagnosis and surgery suffers some inherent disadvantages, i.e., photo bleaching in white light that impairs the possibility for PDD as white light usually is used before the blue light for PDD. Based on spectroscopic observations of urine and the photoactive dye Protoporphyrin IX used in PDD a novel optical system for use with the cystoscope has been devised that solves the problem of green fluorescence from urine. This and the knowledge of photo-bleaching pitfalls in established systems make it possible to perform PDD of bladder tumours in the OPD and to improve PDD in the OR.

Fiber optic attraction of urinary stones during laser lithotripsy has been previously observed, and this phenomenon may potentially be exploited to pull stones inside the urinary tract without mechanical grasping tools, thus saving the urologist valuable time and space in the ureteroscope’s single working channel. In this study, Thulium fiber laser (TFL) high-pulse-rate/low-pulse-energy operation and Holmium:YAG low-pulse-rate/high-pulse-energy operation are compared for fiber optic “suctioning” of Plaster-of-Paris stone phantoms. A TFL with wavelength of 1908 nm, pulse energy of 35 mJ, pulse duration of 500 μs, and pulse rate of 10-350 Hz, and Holmium laser with wavelength of 2120 nm, pulse energy of 35-360 mJ, pulse duration of 300 μs, and pulse rate of 20 Hz were tested using 270-μm-core fibers. A peak “pull” speed of ~ 2.5 mm/s was measured for both TFL (35 mJ and 150-250 Hz) and Holmium laser (210 mJ and 20 Hz). Particle image velocimetry and thermal imaging were used to track water flow for all parameters. Fiber optic suctioning of urinary stone phantoms is feasible for both lasers. However, TFL operation at high-pulse-rates/low-pulse-energies provides faster, smoother stone pulling than Holmium operation at low-pulserates/ high-pulse-energies. After further study, this method may be used to manipulate urinary stones in the clinic.

Q-switched Tm:YAG laser ablation mechanisms on urinary calculi are still unclear to researchers. In this study, dependence of water content in calculus phantom on calculus ablation performance was investigated. White gypsum cement was used as a calculus phantom model. The calculus phantoms were ablated at single pulse and contact mode in three different conditions: dry calculus in air, wet calculus in air, and wet calculus in water. Ablation volume was obtained on average 0.006, 0.008, and 0.008 mm³ in dry calculus in air, wet calculus in air, and wet calculus in water groups, respectively. There were three proposed ablation mechanisms that could explain the effect of water content in calculus phantom on calculus ablation performance, including shock wave due to bubble collapse, spallation, and microexplosion. Shock wave generation due to bubble collapse in wet calculus in water condition had negligible effect on calculus ablation as captured by a needle hydrophone and cannot be a primary mechanism for calculus ablation in this study. Increased absorption coefficient of wet calculus can cause stronger spallation process compared with that caused by dry calculus; and as a result, higher calculus ablation was observed in both wet calculus in air and wet calculus in water. Vaporization of interstitial water in porous calculus phantom can also help enhance calculus ablation efficiency. There were some limitations in this study including use of small sample size and lack of employing real urinary calculus, which should be addressed in future experiment.

The Thulium fiber laser (TFL) has recently been proposed as an alternative to the Holmium:YAG laser for lithotripsy. The TFL’s Gaussian spatial beam profile provides higher power transmission through smaller optical fibers with reduced proximal fiber tip damage, and improved saline irrigation and flexibility through the ureteroscope. However, distal fiber tip damage may still occur during stone fragmentation, resulting in disposal of the entire fiber after the procedure. A novel design for a short, detachable, distal fiber tip that can fit into an ureteroscope’s working channel is proposed. A prototype, twist-lock, spring-loaded mechanism was constructed using micromachining methods, mating a 150-μm-core trunk fiber to 300-μm-core fiber tip. Optical transmission measuring 80% was observed using a 30 mJ pulse energy and 500 μs pulse duration. Ex vivo human calcium oxalate monohydrate urinary stones were vaporized at an average rate of 187 μg/s using 20 Hz modulated 50% duty cycle five pulse packets. The highest stone ablation rates corresponded to highest fiber tip degradation, thus providing motivation for use of detachable and disposable distal fiber tips during lithotripsy. The 1-mm-outer-diameter prototype functioned comparable to previously tested tapered fiber tips, and the novel design shows promise for potential integration into miniaturized flexible ureteroscopes.

OBJECTIVES: We tested the hypothesis that retropulsion varies with stone size. METHODS: Stone phantoms of uniform cube dimensions were constructed and irradiated with Ho:YAG energy (0.5 J - 3.5 J). Displacement was measured. RESULTS: At any given pulse energy, retropulsion decreased as stone size increased, p<0.05. At any given stone size, retropulsion increased as pulse energy increased, p<0.05. CONCLUSIONS: A strategy of low pulse energy at high repetition rate is appropriate for ureteral stones. For larger bladder and renal stones, retropulsion is minimal even with high pulse energies. More study is warranted.

Introduction: Different laser-systems are currently used for stone fragmentation in the upper urinary tract. The aim of our study was to evaluate probe velocity and displacement, retropulsion and fragmentation characteristics two novel devices the electromechanically driven EMS LithoBreaker® (EMS Medical), and of the CO2 cartridge driven LMA StoneBreaker® (Cook Urological) in vitro test models.Testing of the LithoBreaker® included additionally two different cushion guides (harder, softer) to assess the effect of the damper properties on the impulse characteristics. Patients and methods: Maximum probe velocities and displacements were measured using high-speed photography at a resolution of 100.000 frames per second. Repulsion testing was conducted through a 7.5 Fr ureteroscope in an underwater set-up. The probes were projected against a non-frangible led mass placed in a 15 Fr horizontally mounted silicone tube as an in-vitro model of the ureter. Repulsion was determined by measuring the distance the lead mass (0.98g) was displaced. Fragmentation efficiency was assessed by measuring the number of single shots required to break Bego Stone phantoms hard (15:3) and soft (15:6) with an average size of 7.5 mm x 5.5 mm placed on a metal mesh (edge length 3.15mm) into < 3 mm fragments. Mean and standard deviation were computed for all groups and statistical analysis was performed (student’s t-test). Results: The StoneBreaker® yielded the highest velocity of 22.0 ± 1.9 m/sec. followed by the LithoBreaker® assembled with the hard cushion guide of 14.2 ± 0.5 m/sec and the soft probe guide of 11.5 ±0.5 m/sec. accordingly. The maximum probe displacement for the StoneBreaker® was 1.04 mm and for the LithoBreaker® 0.9 mm and 1.1 mm (hard versus soft cushion guide). Repulsion produced using the 1mm probes showed no statistical differences between the devices. Using the 2mm probes, the hardness of the damper used significantly changed the repulsion behaviour of the LithoBreaker®. Using the 1mm probe, the amount of single shots for fragmentation of soft Bego Stones was significantly higher for the LithoBreaker® with soft cushion guide: mean 31.5 ± 11.31 and hard cushion guide: mean 21.5 ± 5.29 compared to the StoneBreaker®: mean 11.2 ± 2.65. Fragmentation efficiency for the hard Bego Stones showed similar statistically significant results. Conclusion: The electromechanic LithoBreaker® and the pneumatic Stonebreaker® were shown to be effective in cracking stone phantoms with relatively low number of pulses. Fragmentation characteristics improved substantially with the higher hardness of the cushion support higher velocity equals higher fragmentation performance of the LithoBreaker®. Repulsion produced were at comparable levels. More testing is required to more detailed information on impulse frequency and capacity for stone clearance time to be used in clinical practice.

Prostate cancer (PCa) is the second most common cancer in US men. Metastasis is the final step of tumor progression and remains the primary cause of PCa death. Hence preclinical, orthotopic models of PCa metastasis are necessary to develop new therapeutics against metastatic disease. Yet unlike irrelevant subcutaneous tumor models, the deployment of orthotopic models of cancer metastasis in drug research and development is limited by the inability to longitudinally monitor cancer progression/regression in response to administration of experimental pharmaceuticals. Recently, a nearinfrared fluorescent protein (iRFP) was created for deeper imaging [1]. Imaging prostate tumor growth and lymph node metastasis in nude mice therefore becomes possible using this new fluorescent gene reporter. In this study, we first developed an intensified CCD (ICCD)-based iRFP fluorescence imaging device. Then human PCa PC3 cell lines expressing iRFP gene reporter were orthotopically implanted in male Nu/Nu mice at 8-10 weeks old. After 6-10 weeks, in vivo, in situ and ex vivo fluorescence imaging was performed. In vivo iRFP fluorescence imaging showed that the detected fluorescence concentrated at the prostate and became stronger over time, indicating the growth of implanted PCa. Fluorescence was non-invasively detected at locations of prostate-draining lymph nodes as early as 5 weeks post implantation, indicating the metastasis to lymph nodes. In situ and ex vivo fluorescence imaging demonstrated that the detected signals from PCa and lymph nodes were correlated with cancer positive status of tissues as assessed through standard pathology.

In recent years, Raman spectroscopy has emerged as a potentially viable tool for automated cancer diagnostics. However, due to the complexity of the signal obtained from a tissue, most of the studies have been confined to statistical analysis of the spectra with principal component analysis being most often reported as the analysis of choice. These types of analyses are sensitive to modification of the Raman spectra due to tissue processing. The study presented here addresses the modifications of the Raman spectra obtained from prostate tissue histopathological slides due to the tissue treatment and its influence on the automated cancer diagnostics via Raman spectroscopy.

Cybesin, a smart contrast agent to target cancer cells, was investigated using a near infrared (NIR) spectral polarization imaging technique for prostate cancer detection. The approach relies on applying a contrast agent that can target cancer cells. Cybesin, as a small ICG-derivative dye-peptide, emit fluorescence between 750 nm and 900 nm, which is in the “tissue optical window”. Cybesin was reported targeting the over-expressed bombesin receptors in cancer cells in animal model and the human prostate cancers over-expressing bombesin receptors. The NIR spectral polarization imaging study reported here demonstrated that Cybesin can be used as a smart optical biomarker and as a prostate cancer receptor targeted contrast agent.

It has been proposed a spectral model to evaluate the biochemical differences between prostate carcinoma and benign fragments using dispersive Raman spectroscopy. We have examined 51 prostate fragments from surgically removed PrCa; each fragment was snap-frozen and stored (-80°C) prior spectral analysis. Raman spectrum was measured using a Raman spectrometer (830 nm excitation) coupled to a fiber-optic probe. Integration time and laser power were set to 50 s and 300 mW, respectively. It has been collected triplicate spectra from each fragment (total 153 spectra). Some samples exhibited a strong fluorescence, which was removed by a 7th order polynomial fitting. It has been developed a spectral model based on the least-squares fitting of the spectra of pure biochemicals (actin, collagen, elastin, carotene, glycogen, phosphatidylcholine, hemoglobin, and water) with the spectra of tissues, where the fitting parameters are the relative contribution of the compounds to the tissue spectrum. The spectra (600-1800 cm^-1 range) are dominated by bands of proteins; it has been found a small difference in the mean spectra of PrCa compared to the benign tissue, mainly in the 1000-1400 cm^-1 region, indicating similar biochemical constitution. The spectral fitting model revealed that elastin and phosphatidylcholine were increased in PrCa, whereas blood and water were reduced in malignant lesions (p < 0.05). A discrimination of PrCa from benign tissue using Mahalanobis distance applied to the contribution of elastin, hemoglobin and phosphatidylcholine resulted in sensitivity of 72% and specificity of 70%.

Gathering vibrational data from the human middle ear is quite difficult. To this date the well-known acoustic probe is used to estimate audiometric parameters, e.g. otoacoustic emissions, wideband reflectance and the measurement of the stapedius reflex. An acoustic probe contains at least one microphone and one loudspeaker. The acoustic parameter determination of the ear canal is essential for the comparability of test-retest measurement situations. Compared to acoustic tubes, the ear canal wall cannot be described as a sound hard boundary. Sound energy is partly absorbed by the ear canal wall. In addition the ear canal features a complex geometric shape (Stinson and Lawton¹). Those conditions are one reason for the inter individual variability in input impedance measurement data of the tympanic membrane. The method of Laser-Doppler-Vibrometry is well described in literature. Using this method, the surface velocity of vibrating bodies can be determined contact-free. Conventional Laser-Doppler-Systems (LDS) for auditory research are mounted on a surgical microscope. Assuming a free line of view to the ear drum, the handling of those laser-systems is complicated. We introduce the concept of a miniaturized vibrometer which is supposed to be applied directly in the ear canal for contact-free measurement of the tympanic membrane surface vibration. The proposed interferometer is based on a Fabry-Perot etalon with a DFB laser diode as light source. The fiber-based Fabry-Perot-interferometer is characterized by a reduced size, compared to e.g. Michelson-, or Mach-Zehnder-Systems. For the determination of the phase difference in the interferometer, a phase generated carrier was used. To fit the sensor head in the ear canal, the required shape of the probe was generated by means of the geometrical data of 70 ear molds. The suggested prototype is built up by a singlemode optical fiber with a GRIN-lens, acting as a fiber collimator. The probe has a diameter of 1.8 mm and a length of 5 mm.

Using data from our previously described otoscope¹ that uses 1310 nm phase-sensitive spectral domain optical coherence tomography (PS-SDOCT), we demonstrate a software technique for improving the signal-to-noise (SNR). This method is a software post-processing algorithm applicable to generic PS-SDOCT data describing phase versus time at a specific depth position. By sub-sampling the time trace and shifting the phase of the subsamples to maximize their correlation, the subsamples can be coherently averaged, which increases the SNR.

Aim: To develop a bilateral vestibulopathy animal model induced by gentamicin using RS rat and to see the effect of LLLT on this bilateral vestibulopathy model. Method: RS rats were divided into 3 groups, control group (C), laser group (L), and histology group (H). All animals in the 3 groups received gentamicin (GM) 110 mg/kg, intravenously once daily for 3 days. The animals underwent sinusoidal oscillation about a vertical axis before the GM injection, 1, 3, and 7 days post injections. Transcanal low level laser therapy (LLLT) was irradiated to left ear canal for 7 days, starting 1 day post the GM injection. The H group animals were irradiated into the left ear of L group for 3 days, starting 1 day post GM injections for 3 days. C and L groups were sacrifice on 9^th day and H group was sacrificed on 7^th day. Results: The gain of the C group was significantly decreased in 3 and 7 days. The gain of the right ear of L group was decreased significantly in 3 and 7 days. The gain of left ear of L group was decreased in 3 days post LLLT but the decreased gain was improved significantly comparing to the level of 7 days gain of right ear and it was much closer to the pre-GM level. The average number of cells in cupula of H group after laser treatment for 3 days was significantly lower in non laser treated right ear comparing to the laser treated left ear and ears of the normal rats. Conclusion: The present study demonstrated that LLLT restores vestibular function and vestibular hair cells in rats post gentamicin induced ototoxic damage. LLLT may have clinical implications in the treatment of various vestibular dysfunction. Further studies are essential to verify the exact mechanisms and the most effective application of LLLT to rescue vestibular dysfunction.

We describe a novel dual-functional optical coherence tomography (OCT) system with both a 3-D OCT real time scanner and a fiber probe using a sapphire ball lens for imaging and sensing the critical structures of the temporal bone. To prevent injury to facial nerve, 3-D visualization links anatomic landmarks to 3-D map of critical intracochlear structures. We used a graphics processing unit to boost the computing and 3-D rendering performance of swept source OCT. Both the intracochlear structures and facial nerve trunk of cadaveric human temporal bones are clearly identified with 3-D OCT volumetric rendering.

A major reason we can perceive faint sounds and communicate in noisy environments is that the outer hair cells of the organ of Corti enhance the sound-evoked motions inside the cochlea. To understand how the organ of Corti works, we have built and tested the phase-sensitive Fourier domain optical coherence tomography (PSFDOCT) system. This system has key advantages over our previous time domain OCT system [1]. The PSFDOCT system has better signal to noise and simultaneously acquires vibration data from all points along the optical-axis [2]. Feasibility of this system to measure in vitro cochlear vibrations in the apex was demonstrated earlier [3]. In this study, we measure the in vivo vibrations of the organ of Corti via round window in live anaesthetized guinea pigs using PSFDOCT. This region of the guinea pig cochlea responds to very high frequencies (10 - 40 kHz). The current vibration noise floor for native organ of Corti tissue is 0.03 nm in this frequency range. Sound-induced vibrations of the stapes, which delivers input to the cochlea, are also measured. The measured vibrations of the organ of Corti demonstrate non-linear compression and active amplification characteristic of sensitive mammalian cochlea.

Objective: To investigate the concordance between optical images obtained with high-resolution microendoscopy (HRME) and conventional histopathology for ex vivo cholesteatoma specimens and surrounding middle ear epithelium. Methods: After resection of cholesteatoma and surrounding middle ear epithelium from surgical patients, tissues were stained with a contrast agent, proflavine, and the HRME fiberoptic scope was placed directly on each tissue specimen. 4- 10 short movie clips were recorded for both the cholesteatoma and surrounding middle ear epithelium specimens. The imaged areas were sent for standard histopathology, and the stained specimens were correlated with the HRME images. IRB approval was obtained, and each patient was consented for the study. Results: Ten cholesteatoma specimens and 9 middle ear specimens were collected from 10 patients. In each case, cholesteatoma was easily discriminated from normal middle ear epithelium by its hyperfluorescence and loss of cellular detail. Qualitative analysis for concordance between HRME images and histological images from the same surgical specimen yielded a strong correlation between imaging modalities. Conclusions: Keratinizing cholesteatoma and surrounding middle ear epithelium have distinct imaging characteristics. Loss of cellular detail and hyperfluorescence with proflavine are the hallmark characteristics of cholesteatoma which allow for differentiation from normal middle ear epithelium. Real-time optical imaging can potentially improve the results of otologic surgery by allowing for extirpation of cholesteatomas while eliminating residual disease. We anticipate performing an in vivo study to test this hypothesis.

The application of photonics to manipulate and stimulate neurons and to study neural networks has gained momentum over the last decade. Two general methods have been used: the genetic expression of light or temperature sensitive ion channels in the plasma membrane of neurons (Optogenetics and Thermogenetics) and the direct stimulation of neurons using infrared radiation (Infrared Neural Stimulation, INS). Both approaches have their strengths and challenges, which are well understood with a profound understanding of the light tissue interaction(s). This paper compares the opportunities of the methods for the use in cochlear prostheses. Ample data are already available on the stimulation of the cochlea with INS. The data show that the stimulation is selective, feasible at rates that would be sufficient to encode acoustic information and may be beneficial over conventional pulsed electrical stimulation. A third approach, using lasers in stress confinement to generate pressure waves and to stimulate the functional cochlea mechanically will also be discussed.

High-speed digital videoendoscopy system is emerging as a new clinical tool for voice assessment. The system can acquire images of the vibrating vocal folds with simultaneous recording of voice data from the patient. The laryngeal image-based analysis has been proven valuable for objective and quantitative assessment of voice kinematics in health and disease, and meanwhile, acoustic analysis of voice data could assist in the study of phonatory characteristics and reveal useful information related to laryngeal pathophysiology. Contrast to the hardware acquisition systems, the development of effective software for handling such massive visual/sound data has lagged behind. In this paper, a software system is designed to process the laryngeal image sequences and perform image-based analyses as well as acoustic analyses. Our software contains following modules: (1) Import and view Module - to read AVI video data and sound data (wave file), edit/compile and save selected data, make image montages using DirectShow technology and display the acoustic waveform using DirectSound technology; (2) Image Process Module – to perform frame-by-frame image segmentation to delineate the glottis, to extract the GAW and bilateral vocal fold displacements; (3) Image Analysis Module – to adopt Nyquist plot displays that involves the Hilbert transform based analysis of GAW, and to provide instantaneous frequency and amplitude distributions; (4) Acoustic Analysis Module – to perform Fast Fourier Transform (FFT) and Spectrogram analyses of the imported sound data, to display the plot of the sound data and provide instantaneous frequency and amplitude distributions and Nyqiust plot and (5) Dual GAW and sound wave display module. Upon rigorous testing of this software using clinical data samples we demonstrate the applications of the software to the study of dynamic characteristics of the glottis, which may correlate with voice quality and health condition.

Acquired subglottic stenosis is a narrowing of the airway caused by prolonged endotracheal intubation. Currently, there are no non-invasive means to diagnose the disease. A previous study by this same group introduced optical coherence tomography (OCT) as a means of monitoring the progression of stenosis. The aim of the current study was to qualitatively and quantitatively analyze OCT images obtained from a subglottic stenosis model of the rabbit airway. 15 rabbits were used throughout the study, and a MEMs based OCT probe was utilized. The OCT images obtained were analyzed using a free software program, 3D Slicer. The region of scarred tissue was grown out and measured quantitatively. This study demonstrated the feasibility of using a program to quantify the progression of scarring in OCT images, in addition to qualitatively correlating between histology, endoscopic, and OCT images. Future works may include utilization of a long-range probe and use of a pressure necrosis model to better emulate the actual onset of neonatal subglottic stenosis.

Divided-pupil line-scanning confocal microscopy (DPLSCM) can provide a simple and low-cost approach for imaging of human tissues with pathology-like nuclear and cellular detail. Using results from a multidimensional numerical model of DPLSCM, we found optimal pupil configurations for improved axial sectioning, as well as control of speckle noise in the case of reflectance imaging. The modeling results guided the design and construction of a simple (10 component) microscope, packaged within the footprint of an iPhone, and capable of cellular resolution. We present the optical design with experimental video-images of in-vivo human tissues.

Late oral radiation toxicity is a common condition occurring in a considerable percentage of head and neck cancer patients after radiation therapy which reduces their quality of life. The current examination of these patients is based on a visual inspection of the surface of the oral cavity; however, it is well known that many of the complications start in the subsurface layers before any superficial manifestation. Considering the currently suboptimal examination techniques, we address this clinical problem by using optical coherence tomography (OCT) to monitor the subsurface oral layers with micron-scale resolution images. A spectral-domain OCT system and a specialized oral imaging probe were designed and built for a clinical study to image late oral radiation toxicity patients. In addition to providing qualitative 2D and 3D images of the subsurface oral layers, quantitative metrics were developed to assess the back-scattering and thickness properties of different layers. Metric derivations are explained and preliminary results from late radiation toxicity patients and healthy volunteers are presented and discussed.

It is generally accepted that oral cancer arises in the presence of oral precancerous lesions. However, the clinical courses of these lesions are quite unpredictable, and a fundamental enigma remains that when and how these lesions turn to malignant growth. Characterization of these potentially malignant lesions is thus important and could serve as early indicators of this neoplastic transformation process, potentially facilitates the treatment outcome and improves the survival rate. Higher harmonic generation microscope (HGM), providing images with a <500nm lateral resolution at a 300μm penetration depth without leaving photodamages in the tissues, was used for this purpose. Oral cavity biopsies were obtained from 18 patients with clinical suspected oral precancerous lesions scheduled for surgical biopsy. HGM images were compared with histological images to determine the results. By visualization of subtle cellular and morphological changes, the preliminary result of this HGM image discloses excellent consistency with traditional histolopathology studies, without the need for fixation, sectioning and staining. More specifically speaking, the keratin thickness was found to be increased comparing with normal adjacent controls. In some cases, variations in cell size, nuclear size and increased nuclear/cytoplasmic ratio, and increased size of nucleoli were identified, indicating different stages of malignant transformation. These results together indicated that HGM provides the capability to characterize features of oral precancerous lesions as well as oral cancer progression, and holds the greatest potential as an ideal tool for clinical screening and surveillance of suspicious oral lesions.

Optical interferometry using Fourier domain OCT (FD-OCT) can image structures using vibration as the contrast mechanism. An A-scan measurement of light reflected from a tissue at a position x,y yields an intensity spectrum, I(λ), which is sequentially acquired over a short time period, 88 ms, to yield 1000 spectra at equally spaced time points, I(λ,t), while an audio frequency of sound pressure is imparted to the tissue to induce vibration. Analysis converts I(λ,t) into displacement Δz(z,t) of tissue structures along the z axis. Fourier analysis converts Δz(t) at each z into a vibration spectrum, and the amplitude of vibration at the driving audio frequency is specified. This process is repeated for each x position at a given y, to yield an image of vibration amplitude, A(z,x) [nm]. Hence, vibration amplitude becomes the contrast mechanism. This method is not a new idea, but a derivative of Doppler OCT. This report simply provides a concise algorithm in MATLAB for students wishing to implement vibration measurements using FD-OCT. An example shows the vibration of the ossicles of the middle ear viewed through the intact tympanic membrane.

Melanoma of the head and neck and its treatment are complex issues. The behavior of head and neck melanoma is aggressive, and it has an overall poorer prognosis than of other skin sites. The goal of this research was to determine if melanoma manipulation could enhance penetration of cancer cells from the primary tumor into the circulatory system. Nude mice were inoculated with melanoma cells in the mouse ear. Blood vessels were monitored for the presence of circulating tumor cells (CTCs) using in vivo photoacoustic (PA) flow cytometry (PAFC). The implanted tumor underwent compression, incisional biopsy, or surgical excision, and the release of CTCs was monitored using in vivo PAFC in real time. We discovered that some medical procedures, like compression of an implanted tumor of a mouse model or an incisional biopsy, may either initiate CTC release in the blood which previously contained no CTCs or dramatically increased (10-30–fold) CTC counts above the initially recorded level. These findings indicate that some intervention (such as palpation during physical exam, or incisional biopsy) can potentially enhance penetration of cancer cells from a primary tumor into the blood circulation, which may increase the risk of metastases. Our results warn oncologists of at least some precaution during physical examination, careful surgery strategy.

Introduction: The need for reduction of post-tonsillectomy hemorrhage has led to promotion of tonsillotomy techniques for tonsil tissue reduction in obstructive tonsillar hypertrophy. A first study compares ablative tissue effects using 1470nm diode laser and CO2-laser for tonsillotomy in an intraindividual design. A number of different laser systems have been used for volume reduction of hyperplastic nasal turbinates. The aim of a 2nd clinical feasibility study was to show the coagulative and tissue reducing effects using a novel Tm: fiber laser system emitting at λ = 1940 nm Patients and methods: First 21 children aged 3 -13 years (mean age 6.3 years) underwent laser tonsillotomy for obstructive tonsillar hypertrophy in this double blind, prospective, randomized, clinical feasibility trial. In each case, tonsillotomy was performed using fibre guided 1470nm diode laser (contact mode, 15 W power) on the one side and CO2-laser (12 W power) on the other side. An independent physician documented clinical presentation and patients’ symptoms preoperatively and on day 1, 3, 7, 14 and 21 postoperatively using standardized questionnaire including VAS (was ist das) for each side separately. The 2^nd clinical feasibility trial included 11 patients suffering from hyperplastic inferior nasal turbinates, who were therapy-refractory to conservative medical treatment. The obstructive nasal cavity was treated using the 1940 nm Tm: fiber laser at < 5 W output power. The treatment was performed in non-contact mode under endoscopic control. Patients ’ symptoms were documented both preoperatively and on days 1 – 3 and 28 postoperatively using a non-validated questionnaire. Additionally, an endoscopic examination was performed. Results: Mean duration of single tonsillotomy operative treatment was 2.7 min using 1470nm laser and 4.9 min using CO2 laser respectively. Intraoperative bleeding and the frequency of bipolar forceps use for intraoperative bleeding control was significantly less pronounced using the 1470nm diode laser system. There was no difference in postoperative pain scores between the CO2-laser treated and the 1470nm fibre guided diode laser treated side. No infections, hemorrhages or other complications occurred in the course of the three weeks postoperative period. In the turbinate study, none of the patients showed infections, and no hemorrhages or other complications occurred intraor postoperatively.The mean laser activation time was extremely short being 28.0 ± 8.5 s. In conjunction with a low power setting (median, 3 W; mean ± standard deviation, 3.3 ± 1.1 W), a low energy of 90.2 ± 37.8 J was applied. A significant reduction in nasal obstruction could be documented in all patients on day 28 postoperatively. Evaluation, as assessed preoperatively and 4 weeks postoperatively, showed significant subjective improvements. Conclusion: A fiber-guided 1470nm diode laser system offers an efficient and safe method for tonsillotomy as treatment of obstructive tonsillar hypertrophy. Compared to our standard practice with CO2- laser, 1470nm laser application provides comparable tissue ablation effects with less intraoperative bleeding and shorter operation time. The treatment of hyperplastic inferior turbinates using a 1940 nm Tm: fiber laser provides sufficient tissue reduction in a short operation time using low total energy. Patients described a significant improvement in nasal breathing postoperatively.

This paper describes the development and implementation of 3 μm lasers for myringotomy and microsurgery. Two different lasers were investigated. The first, an Er-doped, CW zirconate glass fiber laser optically pumped by a 970 nm diode laser, emitted > 1 W of CW power at 2.76 μm with concomitant green incoherent emission that served as a convenient visible illumination beam. The second, a 1 W CW Er:YAG solid-state laser also optically pumped by a 970 nm diode laser, emitted > 1 W of CW power at 2.94 μm, coincident with the strongest infrared water absorption peak. Running CW, both lasers are expected to avoid the loud acoustical shocks associated with pulsed lasers. Myringotomies were carried out with the Er:YAG laser on anaesthetized guinea pigs and the effects of the laser were documented. Laser ablated samples of tympanic membrane, soft tissue and bone were histologically examined. Histology results indicated that the CW Er:YAG laser is a potential candidate for a new myringotomy tool and possibly for otologic microsurgery, but deliverable power levels need to be increased to the 2 W (or higher) level. This work was funded under NIH SBIR Grant No. 5R44DC004899.

Background and Purpose: Combination therapy of interstitial photodynamic therapy (iPDT) with Cetuximab to attain symptomatic control of recurrent head and neck cancer.

Methods: Two patients with Unresectable recurrent Head and Neck SCC were treated with iPDT alone and iPDT and cetuximab. Treatments were administered in an outpatient setting. A single dose of Photofrin at 2 mg per kilogram of body weight was administered intravenously two days prior to laser illumination. The iPDT was accomplished by delivering 630-nm laser light through two laser fibers with 2.5 and 5 cm long diffusive ends. Light irradiance of 400 mW/cm for 250 seconds was used to deliver a total of 100 J/cm, during the iPDT. Light applications were conducted, twice, at 3-4 days interval. One of the patients was treated with cetuximab along with iPDT.

Results: Near total resolution of tumor was observed in the patient treated with iPDT and cetuximab, and partial resolution was seen in the patient treated with iPDT alone.

Conclusion: Interstitial photodynamic therapy may be used to treat patients with recurrent unresectable head and neck cancer. The combination of iPDT with Cetuximab has the potential to improve tumor response in the patient population for whom there is no effective therapies. This observation merits further studies.

In this paper, we conduct a phantom study for modeling the autofluorescence (AF) properties of tissue. A combined optical coherence tomography (OCT) and AF imaging system is proposed to measure the strength of the AF signal in terms of the scattering layer thickness and concentration. The combined AF-OCT system is capable of estimating the AF loss due to scattering in the epithelium using the thickness and scattering concentration calculated from the co-registered OCT images. We define a correction factor to account for scattering losses in the epithelium and calculate a scatteringcorrected AF signal. We believe the scattering-corrected AF will reduce the diagnostic false-positives rate in the early detection of airway lesions due to confounding factors such as increased epithelial thickness and inflammations.

Introduction: A recent ex-vivo study using micro-CT in patients with chronic obstructive pulmonary disease (COPD) showed that narrowing and disappearance of small conducting airways precedes the onset of emphysematous destruction in COPD. Until recently, the airway remodeling process could not be studied in detail in-vivo. In this study, we investigated the repeatability of navigating an Optical Coherence Tomography (OCT) catheter to image the same airways in smokers with and without COPD. Method: OCT imaging was performed by inserting the catheter through a sub-segmental airway to a small bronchiole. Three-dimensional OCT imaging of 5 cm of airway segments was obtained. The catheter was removed and reinsertion into the same airway was attempted. The number of airway generations and quantitative measurements of the airway wall area were investigated. Results: Sixty-three airways in 30 subjects were analyzed. Repeated insertion into the same airway was observed at 53.8 %, 92.3% and 70.8% of the time in the upper, middle and lower lobes respectively. The percentage differences of paired measurements of airway wall area between matched and unmatched airways in bronchioles were 5.8 ± 4.6 % and 7.3 ± 5.4 % respectively Conclusions: Repeated OCT imaging of airways is possible in the majority of cases except in the upper lobes. For airways that are not completely matched, some of the airway segments can still be used for comparison by careful alignment of the airway. OCT may be a useful method to study the remodeling process in small airways and the effect of therapeutic intervention.

Lung cancer is the leading cause of cancer related death. Macroscopic imaging techniques such as computed tomography are highly sensitivity at detecting small, ≤ 2cm, peripheral pulmonary lesions (PPLs) in the lung but lack the specificity necessary for diagnosis. Bronchoscopy is a procedure routinely performed to diagnose PPLs but is hindered with a low diagnostic yield due to challenging lesion localization. We have developed a flexible transbronchial optical frequency domain imaging (TB-OFDI) catheter that functions as a ‘smart needle’ to confirm the needle placement within the target lesion prior to biopsy. The TB-OFDI smart needle consists of a flexible and removable OFDI catheter that operates within a 21-gauge transbronchial needle aspiration (TBNA) needle. The OFDI catheter can be easily removed from the needle to facilitate subsequent aspiration or biopsy acquisition. The OFDI imaging core consists of an angled-polished ball lens with a spot size of 25 μm at a working distance of 160 μm from the catheter sheath. The ball-lens was designed to have an ellipsoid shape in order to compensate for the astigmatism caused by encasing the optics within a protective sheath. Transbronchial imaging of inflated excised swine lung parenchyma with the TB-OFDI smart needle yielded clear images of alveoli. In-vivo transbronchial imaging was also performed on three swine with artificial lesions injected transthoracially. Our results suggest that the TB-OFDI smart needle may be a useful tool for guiding biopsy acquisition to increase the diagnostic yield of PPLs.

We have been investigating ultrahigh resolution optical coherence tomography (UHR-OCT) imaging of lung tissues using fiber based super continuum (SC) sources. The high power, low-noise, Gaussian shaped SC generated with ultrashort pulses and optical fibers at several wavelength regions were used as the broadband light sources for UHROCT. Since the lung consists of tiny alveoli which are separeted by thin wall, the UHR-OCT is supposed to be effective for lung imaging. The normal and diseased lung tissues were observed without invasive procedures to the lung itself. The clear images of alveoli were observed with index matching effect by saline. In this work, we investigated the three-dimensional UHR-OCT imaging of lung structure. The lungs of rats inflated with 10% formalin at 5 cmH₂O, 15 cmH₂O, and 20 cmH₂O pressure were prepared as the sample for investigation of size and shape of the lung structure. These samples were fixed with 10% formalin. The interalveolar septa, thin walls separating the alveoli, were clearly observed. The difference of size and shape of alveoli and thier three-dimensional network was clearly observed from the UHR-OCT images. The clear images of alveoli were observed with index matching effect of 10% formalin. We investigated the wavelength dependence of 3D UHR-OCT image of lung structure at 800 nm, 1060 nm, and 1700 nm wavelength regions. The 3D UHR-OCT images of structure of rat lung were clearly observed in all wavelength regions and wavelength dependence of imaging was discussed.

New optical techniques have the potential to fill the gap between radiological and microscopic approaches to assess the lung's internal structure. Since its quantitative assessment requires unbiased sampling and measurement principles, imaging of the whole lung with sufficient resolution for visualizing details is important. To address this request, we applied scanning laser optical tomography (SLOT) for the three dimensional imaging of mouse lung ex vivo. SLOT is a highly efficient flourescence and transmission microscopy technique allowing for 3D imaging of specimen of sizes up to several millimeters. Previously fixed lung lobes and whole lungs were optically cleared and subsequently imaged with SLOT while making use of intrinsic contrast mechanisms like absorption and autofluorescence. Imaging of airways, blood vessels and parenchyma is demonstrated. Volumetric SLOT datasets of the lung's internal structure can be analyzed in any preferred planar orientation. Moreover, the sample preparation preserves microscopic structure of the lung and allows for subsequent correlative histologic studies. In summary, SLOT is a useful technique to visualize and survey the internal structure of mouse lung at different scales and with various contrast mechanisms. Potential applications of SLOT in lung research are e.g. quantitative phenotype analysis of mouse models of human lung disease in combination with stereological methods.

Tuberculosis, caused by Mycobacterium tuberculosis (Mtb), currently affects roughly one-third of the world’s population. Drug resistant strains of Mtb decrease the effectiveness of current therapeutics and demand the development of new antimicrobial therapies. In addition, the current vaccine, Bacille Calmette Guérin (BCG), has variable efficacy for disease prevention in different populations. Animal studies are often limited by the need to sacrifice at discrete time points for pathology and tissue homogenization, which greatly reduces spatial and temporal resolution. Optical imaging offers the potential for a minimally-invasive solution to imaging on a macroscopic and microscopic scale, allowing for high resolution study of infection. We have integrated a fluorescence microendoscope into a whole-animal optical imaging system, allowing for simultaneous microscopic and macroscopic imaging of tdTomato expressing BCG in vivo. A 535 nm LED was collimated and launched into a 10,000 element fiber bundle with an outer diameter of 0.66 mm. The fiber bundle can be inserted through an intra-tracheal catheter into the lung of a mouse. Fluorescence emission can either be (1) collected by the bundle and imaged onto the surface of a CCD camera for localized detection or (2) the fluorescence can be imaged by the whole animal imaging system providing macroscopic information. Results from internal localized excitation and external whole body detection indicate the potential for imaging bacterial infections down to 100 colony forming units. This novel imaging technique has the potential to allow for functional studies, enhancing the ability to assess new therapeutic agents.

The development of real-time, label-free imaging techniques has recently attracted research interest for in situ differentiation of cancerous lesions from normal tissues. Molecule-specific intrinsic contrast can arise from label-free imaging techniques such as Coherent Anti-Stokes Raman Scattering (CARS), Two-Photon Excited AutoFluorescence (TPEAF), and Second Harmonic Generation (SHG), which, in combination, would hold the promise of a powerful label-free tool for cancer diagnosis. Among cancer-related deaths, lung carcinoma is the leading cause for both sexes. Although early treatment can increase the survival rate dramatically, lesion detection and precise diagnosis at an early stage is unusual due to its asymptomatic nature and limitations of current diagnostic techniques that make screening difficult. We investigated the potential of using multimodality nonlinear optical microscopy that incorporates CARS, TPEAF, and SHG techniques for differentiation of lung cancer from normal tissue. Cancerous and non-cancerous lung tissue samples from patients were imaged using CARS, TPEAF, and SHG techniques for comparison. These images showed good pathology correlation with hematoxylin and eosin (H and E) stained sections from the same tissue samples. Ongoing work includes imaging at various penetration depths to show three-dimensional morphologies of tumor cell nuclei using CARS, elastin using TPEAF, and collagen using SHG and developing classification algorithms for quantitative feature extraction to enable lung cancer diagnosis. Our results indicate that via real-time morphology analyses, a multimodality nonlinear optical imaging platform potentially offers a powerful minimally-invasive way to differentiate cancer lesions from surrounding non-tumor tissues in vivo for clinical applications.