This PDF file contains the front matter associated with SPIE Proceedings Volume 10038, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

Bladder cancer is one of the most expensive cancers to manage due to frequent recurrences requiring life-long surveillance and treatment. A near-infrared labeled 2-deoxy-d-glucose probe IRDye800CW-DG targeting glucose metabolism pathway has shown to enhance the sensitivity of diagnosing several types of cancers as tested on tumor models not including bladder tumor. This pilot study has explored differential uptake of intravesically administered IRDye800CW-DG in an orthotopic rat bladder tumor model. Twenty-five female Fischer rats were randomly grouped to four conditions: no-tumor-control (n=3), no-tumor-control intravesically instilled with IRDye800CWDG (n=6), rats bearing GFP-labeled AY-27 rat bladder urothelial cell carcinoma cells and washed with saline (n=5), and rats bearing AY-27 tumors and intravesically instilled with IRDye800CW-DG (n=11). Near-infrared fluorescence was measured from the opened bladder wall of anesthetized rat at an excitation wavelength of 750nm and an emission wavelength of 776nm, by using an in-house fluorescence imaging system. There is no statistically significant difference of the peak fluorescence intensity among the no-tumor-control bladders (n=3), the no-tumorcontrol bladders instilled with IRDye800CW-DG (n=6), and the GFP-labeled AY-27 treated bladders washed by saline (n=5). When compared to that of the no-tumor-control bladders instilled with IRDye800CW-DG (n=6), the fluorescence intensity of GFP-labeled AY-27 treated bladders instilled with IRDye800CW-DG and with histology confirmed neoplastic bladder tissue (n=11) was remarkably more intense (3.34 fold of over the former) and was also statistically significant (p<0.0001). The differential uptake of IRDye800CW-DG by the neoplastic urinary bladder tissues suggests the potential for cystoscopy-adaptation to enhance diagnosis and guiding surgical management of flat urinary bladder cancer.

More than 1 million men in the United States undergo a prostate biopsy procedure annually and approximately 200,000 men receive a diagnosis of prostate cancer. 5-10% of these men have to undergo a repeat biopsy due to insufficient tissue sampling. We are studying the utility of a multi-spectral time resolved fluorescence spectroscopy (MS-TRFS) technique for real-time prostate cancer diagnosis. The MS-TRFS imaging setup, which includes a fiberoptic set-up with a 355nm excitation light source coupled with a blue (450nm) aiming beam, was used to image ex-vivo prostatectomy specimen. The prostate tissue from 11 patients was sectioned at 2mm thickness and the fluorescence lifetime information was overlaid spatially for histology and thus, diagnostic co-registration. Initial results show that fluorescence lifetime in the 390±40nm channel, which measures collagen and elastin signatures, is longer for glandular regions than in the stromal regions. Additionally, lifetime in the 452±45nm channel, corresponding to NAD redox state, is longer in the cancerous glandular region in comparison with the normal glandular regions. Current work is focused on developing real-time quantitative algorithms to combine the fluorescence signatures from the two channels for performing prostate cancer diagnosis on biopsies.

Urinary stones harvested from patients were under in-vitro investigation. Fluorescence measurements were performed either by taking images under blue light excitation light as well as measuring excitation-emission-matrixes. Ho:YAG-laser assisted fragmentation was performed in an aquarium set-up to derived fragmentation/dusting rates. FTIR-spectroscopy was used to identify the composition of the stone. Blue light fluorescence excitation resulted in fluorescence emission in different spectral regions spectroscopically proven by EEM-measurement. A correlation with FTIR-spectroscopy is performed. Fragmentation experiments resulted in a dependency to the applied energy/pulse and repetition rate. Using photonic techniques urinary stones could be categorized by their composition. The impact of fluorescence guidance during endoscopic laser lithotripsy will be discussed.

Transurethral resections are commonly used for bladder cancer diagnosis, treatment and follow-up. Cancer staging relies largely on the analysis of muscle in the resections; however, muscle presence is uncertain at the time of the resection. An extemporaneous quality control tool would be of great use to certify the presence of muscle in the resection, and potentially formulate a primo-diagnosis, in order to ensure optimum patient care. Full-field optical coherence tomography (FFOCT) offers a fast and non-destructive method of obtaining images of biological tissues at ultrahigh resolution (1μm in all 3 directions), approaching traditional histological sections. This study aimed to evaluate the potential of FFOCT for the quality control and the primo-diagnosis of transurethral bladder resections. Over 70 transurethral bladder resections were imaged with FFOCT within minutes, shortly after excision, and before histological preparation. Side-by-side comparison with histology allowed to establish reading criteria for the presence of muscle and cancer in particular. Images of 24 specimens were read blindly by three non-pathologists readers: two resident urologists and a junior bio-medical engineer, who were asked to notify the presence of muscle and tumor. Results showed that after appropriate training, 96% accuracy could be obtained on both tumour and muscle detection. FFOCT is a fast and nondestructive imaging technique that provides analysis results concordant with histology. Its implementation as a quality control and primo-diagnosis tool for transurethral bladder resections in the urology suite is feasible and lets envision high value for the patient.

Urinary tract infections(UTI) pose a serious problem for hospital patients accounting for 33% of all hospital acquired(nosocomial) infections with indwelling foley catheters. The presence of an indwelling foley catheter provides a scaffolding for circulating planktonic bacteria to adhere to and to form microbial biofilm communities that would typically be hindered by the body’s innate immune system response. It is these biofilm communities that form on the inner lumen of foley catheters that provide a reservoir of pathogenic bacteria that could dislodge or disperse from the biofilm and infect urethra or bladder mucosal tissue in the urinary tract. Current diagnostic techniques of urine microbiological cultures are lacking in differentiating asymptomatic bacteriuria and symptomatic catheter-associated urinary tract infection(CAUTI) since almost all patients with chronic indwelling catheters are almost universally bacteriuruic. There is an unmet need of a diagnostic tool to assess the difference between the pathogenesis of asymptomatic bacteriuria and CAUTI, specifically at the site of the native biofilm formation. Optical Coherence Tomography(OCT) is an emerging high resolution, minimally invasive tomographic imaging technique that has shown promise in imaging biofilm structures previously in an endoscopic setting of the airway in-vivo and in microfluidic chambers. OCT can be adapted to image various sized biological surfaces and orifices such as airway branches and blood vessels by using a variety of minature endoscopic probes. In this work OCT will be used to image biofilm structure in-vitro on the inner lumen of extravasated critical care patient’s foley catheters. Scanning electron microscopy will be conducted post OCT to confirm the presence of bacterial biofilm in OCT images.

Prostate cancer treatment is shifting from radical to focal therapy. Instant tumor visualization on a microscopic level is crucial for clinical application of focal therapy. Optical coherence tomography (OCT) produces instant tissue visualization on a µm scale and the attenuation of OCT signal as a measure of tissue organization. The objective is to correlate qualitative and quantitative OCT analysis with histopathology. Twenty prostates were analyzed by needle based OCT after radical prostatectomy. For precise correlation, whole mount histology slides were cut through the OCT trajectory. OCT images were classified in eight histological categories. Two reviewers independently performed assessment of the OCT images into these categories. Quantitative attenuation coefficient was used to discriminate stroma and malignant tissue. Sensitivity and specificity for detection of malignancy on OCT was calculated. Visual analyses showed that OCT can reliably differentiate between fat, cystic and regular atrophy and benign glands. Differentiation of benign stroma and inflammation and also malignancy Gleason 3 and 4 is more difficult. Sensitivity and specificity for detection of malignancy on OCT were calculated at 77% and 75%. Quantitative analysis by means of the attenuation coefficient for differentiation between stroma and malignancy showed no significant difference (4.39 mm-1 vs. 5.31 mm-1). Precise correlation of histology and OCT is possible and helps us to understand what we see and measure on OCT. Visual malignancy detection shows reasonable sensitivity and specificity. Our future studies focus on improving discrimination of malignancy with OCT for example by combining an extra imaging modality.

Near-IR laser energy in conjunction with applied tissue cooling is being investigated for thermal remodeling of endopelvic fascia during minimally invasive treatment of female stress urinary incontinence. Previous simulations of light transport, heat transfer, and tissue thermal damage have shown that a transvaginal approach is more feasible than a transurethral approach. However, undesirable thermal insult to vaginal wall was predicted. This study explores whether an optical clearing agent (OCA) can improve optical penetration depth and completely preserve vaginal wall during subsurface treatment of endopelvic fascia. Several OCA mixtures were tested, and 100% glycerol was found to be optimal. Optical transmission studies, optical coherence tomography, reflection spectroscopy, and computer simulations of thermal damage to tissue using glycerol were performed. The OCA produced a 61% increase in optical transmission through porcine vaginal wall at 37 °C after 30 min. Monte Carlo (MC) light transport, heat transfer, and Arrhenius integral thermal damage simulations were performed. MC model showed improved energy deposition in endopelvic fascia using OCA. Without OCA, 62, 37, and 1% of energy was deposited in vaginal wall, endopelvic fascia, and urethral wall, compared with 50, 49, and 1% with OCA. Use of OCA also yielded 0.5 mm increase in treatment depth, allowing potential thermal tissue remodeling at 3 mm depth.

Temperature variations are often monitored by using sensors operating at the site of treatment during Laser-induced Interstitial Thermotherapy (LITT). Currently, temperature measurements during LITT have been performed with thermocouples (TCs). However, TCs could directly absorb laser light and lead to self-heating (resulting in an over-estimation). Fiber Bragg grating (FBG) sensors can instead overcome this limitation of the TCs due to its insensitivity to electromagnetic interference. The aim of the current study was to quantitatively evaluate the FBG temperature sensor with a K-type thermocouple to real-time monitor temperature increase in ex vivo tissue during diffuser-assisted LITT. A 4-W 980-nm laser was employed to deliver optical energy in continuous mode through a 600-µm core-diameter diffusing applicator. A goniometric measurement validated the uniform light distribution in polar and longitudinal directions. The FBG sensor showed a linear relationship (R2 = 0.995) between wavelength shift and temperature change in air and tissue along with a sensitivity of ~ 0.0114 nm/˚C. Regardless of sensor type, the measured temperature increased with irradiation time and applied power but decreased with increasing distance from the diffuser surface. The temperature elevation augmented the degree of thermal coagulation in the tissue during LITT (4.0±0.3-mm at 99˚C after 120-s). The temperature elevation augmented the degree of thermal coagulation in the tissue during LITT s irradiation). The FBG-integrated diffuser was able to monitor the interstitial temperature in tubular tissue (porcine urethra) real-time during laser treatment. However, the thermal coagulation thickness of the porcine urethra was measured to be 1.5 mm that was slightly thicker (~20%) than that of the bovine liver after 4-W 980-nm laser for 48 s. The FBG temperature sensor can be a feasible tool to real-time monitor the temporal development of the temperature during the diffuser-assisted LITT to treat urethral disease.

The Thulium fiber laser (TFL) is being studied as an alternative to Holmium:YAG laser for lithotripsy. The near single mode TFL beam profile enables transmission of higher laser power through smaller optical fibers than possible during Holmium laser lithotripsy. Current free-space coupling of collimated TFL output beam into a disposable silica fiber for ureteroscopy is limited by back-reflected light from the fiber input surface, which may result in laser shutoff or damage, if left unchecked. This study examines whether anti-reflection (AR) coated fibers may sufficiently reduce back-reflected light to prevent laser shutoff, increase fiber optic transmission, and potentially increase laser stone ablation rates as well. Fiber optic transmission and stone ablation studies were conducted comparing uncoated and AR-coated 105- and 200-μm-core fibers. Magnified images of proximal fiber surfaces were taken before and after each trial to examine for AR-coating damage. TFL wavelength of 1908 nm was coupled into silica fibers, with incrementally increasing pulse energy (5-35 mJ), fixed 500-μs pulse duration, and pulse rates of 50-300 Hz. For each pulse rate, 100,000 pulses were also delivered through the fibers to examine for potential damage. Back-reflection at proximal fiber surface was reduced from 3.25% with uncoated fibers to ~ 0.06% with AR coated fibers. For both fiber diameters, output power was stable, and no proximal fiber damage was observed after delivery of 100,000 pulses at 35 mJ, 300 Hz, and 10.5 W average power. There was no significant difference in stone ablation rates between fiber diameters (105 vs. 200 μm) or bare or AR-coated fibers. Laser shutdown was not observed using AR-coated fibers, which reduce back-reflection and improve energy transmission, but do not improve stone ablation rates.

Laser lithotripsy is the preferred application for the destruction of ureteral and kidney stones. Clinically Ho:YAG lasers (λ=2.1μm) are used due to high absorption by water to induce thermomechanical ablation. This study focussed on the investigation of different laser parameters in relation to the stone dusting efficiency. The term dusting was defined when the ablated fragments were d<1mm in diameter while fragmentation is defined to pieces of d> 1mm. The discussion about fragment-size showed advantages like reduced surgery time. Experiments were performed using clinical available Ho:YAG laser energy transferred via a standard fibre (Ø: 365μm) onto phantom calculi (Bego-Stones of different hardness) in a water filled vessel. Dusting can be reached most efficient by using low energy/pulse (approx. 0.5J/pulse) and repetition rate of around 40 Hz. Higher energy/pulse showed strong repulsion and thereby increased mobility, while using lower repetition rates result in longer ablation times. With regard to the hardness of the phantoms it can be derived that on soft calculi or calculi with a very rugged surface dusting can be observed less because the stone breaks into large fragments after a short time of laser application. For hard calculi the ablation process takes a much longer time compared to soft stones. In the following will be shown that dusting and fragmentation process depends not only on the energy/pulse and repetition rate of a Ho:YAG-laser, but also there are differences between Ho:YAG-laser systems according to the dusting efficiency.

The Thulium fiber laser (TFL) is being explored as an alternative to Holmium:YAG laser for lithotripsy. TFL beam profile allows coupling of higher power into smaller fibers than multimode Holmium laser beam, without proximal fiber tip degradation. A smaller fiber provides more space in ureteroscope working channel for increased saline irrigation and allows maximum ureteroscope flexion. However, distal fiber tip burnback increases as fiber diameter decreases. Previous studies utilizing hollow steel sheaths around recessed distal fiber tips reduced fiber burnback, but increased retropulsion. In this study, a “fiber muzzle brake” was tested for reducing fiber burnback and stone retropulsion. TFL lithotripsy studies were performed at 1908 nm, 35 mJ, 500 μs, and 300 Hz using a 100-μm-core fiber. The optimal stainless steel muzzle brake tip tested consisted of a 1-cm-long, 560-μm-OD, 360-μm-ID tube with 275-μm thru hole located 250-μm from the distal end. The fiber tip was recessed a distance of 500 μm. Stone phantom retropulsion, fiber tip burnback, and calcium oxalate stone ablation studies were performed, ex vivo. Small stones with a mass of 40 ± 4 mg and 4-mm-diameter were ablated over a 1.5-mm sieve in 25 ± 4 s (n=10), without distal fiber tip burnback. Reduction in stone phantom retropulsion distance by 50% and 85% was observed when using muzzle brake tips versus 100-μm-core bare fibers and hollow steel tip fibers. The muzzle brake fiber tip provided efficient stone ablation, reduced stone retropulsion, and minimal fiber degradation during TFL lithotripsy.

When treating ureteral calculi, retropulsion can be reduced by using a longer pulse width without compromising fragmentation efficiency (from the studies by David S. Finley et al. and Hyun Wook Kang et al.). In this study, a lab build Ho:YAG laser was used as the laser pulse source, with pulse energy from 0.2J up to 3.0 J, and electrical pump pulse width from 150 us up to 1000 us. The fiber used in the investigation is a 365 μm core diameter fiber, SureFlexTM, Model S-LLF365. Plaster of Paris calculus phantoms were ablated at different energy levels (0.2, 0.5, 1, 2, 3J) and with different number of pulses (1, 3, 10) using different electrical pump pulse width (333, 667, 1000 μs). The dynamics of the recoil action of a calculus phantom was monitored using a high-speed camera with frame rate up to 1 million frame per second (Photron Fastcam SA5); and the laser-induced craters were evaluated with a 3-D digital microscope (Keyence VHX-900F). A design of experiment software (DesignExpert-10, Minneapolis, MN, USA) is used in this study for the best fit of surface response on volume of dusting and retropulsion amplitude. The numerical formulas for the response surfaces of dusting speed and retropulsion amplitude are generated. More detailed investigation on the optimal conditions for dusting of other kinds of stone samples and the fiber size effect will be conducted as a future study.

Background: Following human renal allograft transplant primary graft dysfunction can occur early in the postoperative period as a result of acute tubular necrosis, acute rejection, drug toxicity, and vascular complications. Successful treatment of graft dysfunction requires early detection and accurate diagnosis so that disease-specific medical and/or surgical intervention can be provided promptly. However, current diagnostic methods are not sensitive or specific enough, so that identifying the cause of graft dysfunction is problematic and often delayed. Near-infrared spectroscopy (NIRS) is an established optical method that monitors changes in tissue hemodynamics and oxygenation in real time. We report the feasibility of directly monitoring kidney the kidney in an animal model using NIRS to detect renal ischemia and hypoxia. Methods: In an anesthetized pig, a customized continuous wave spatially resolved (SR) NIRS sensor was fixed directly to the surface of the surgically exposed kidney. Changes in the concentration of oxygenated (O2Hb) deoxygenated (HHb) and total hemoglobin (THb) were monitored before, during and after renal artery clamping and reperfusion, and the resulting fluctuations in chromophore concentration from baseline used to measure variations in renal perfusion and oxygenation. Results: On clamping the renal artery THb and O₂Hb concentrations declined progressively while HHb rose. With reperfusion after releasing the artery clamp O₂Hb and THb rose while HHb fell with all parameters returning to its baseline. This pattern was similar in all three trials. Conclusion: This pilot study indicates that a miniaturized NIRS sensor applied directly to the surface of a kidney in an animal model can detect the onset of renal ischemia and tissue hypoxia. With modification, our NIRS-based method may contribute to early detection of renal vascular complications and graft dysfunction following renal transplant.

The gold standard for the detection of bladder cancer is white light cystoscopy, followed by an invasive biopsy and pathological examination. Tissue pathology is time consuming and often prone to sampling errors. Recently, optical spectroscopy techniques have evolved as promising techniques for the detection of neoplasia. The specific goal of this study is to evaluate the application of combined auto-fluorescence (excited using 378 nm and 445 nm wavelengths) and diffuse reflectance spectroscopy to discriminate normal bladder tissue from tumor at different grades. The fluorescence spectrum at both excitation wavelengths showed an increased spectral intensity in tumors with respect to normal tissues. Reflectance data indicated an increased reflectance in the wavelength range 610 nm - 700 nm for different grades of tumors, compared to normal tissues. The spectral data were further analyzed using principal component analysis for evaluating the sensitivity and specificity for diagnosing tumor. The spectral differences observed between various grades of tumors provides a strong genesis for the future evaluation on a larger patient population to achieve statistical significance. This study indicates that a combined spectroscopic strategy, incorporating fluorescence and reflectance spectroscopy, could improve the capability for diagnosing bladder tumor as well as for differentiating tumors in different grades.

Prostate cancer is the most frequent diagnosed cancers among men. When prostate cancer occurs, the cancer does not result in only one or few localized malignant tumor, but is generally spread within the whole prostate. In order to counteract the very high level of heterogeneities exhibited by prostate tissues, we developed a method for high-resolution co-registration of Raman spectroscopy with prostate cancer diagnosis. Raman spectra were acquired on fresh ex vivo prostate within 2 hours after radical prostatectomy using a multi-wavelength hand-held contact probe. After the measurements, the prostate was reintegrated to the usual pathological workflow: formalin fixated and paraffin embedded (FFPE), and prepared for microscope histopathological analyses. The precise reconstruction of the prostate slice with hematoxylin and eosin (H and E) tissue allows the spatial correlation of the measured area (0.2 mm2) with the correspondent histopathological information, for point-by-point diagnosis determination. The tissue was classified into groups (normal/cancer) and subgroups according to the percentage of benign glands, stroma or cancer. Different machine learning algorithms were tested to classify the spectra with increasing levels of categorization. Preliminary results showed that Raman spectroscopy is capable of detecting prostate cancer with an accuracy >90%. In addition, high percentages of stroma (vs. glands) have been correlated with spectral signature of collagen, which is the main constituent of extracellular matrix.

In vivo single-fiber reflectance spectroscopy (SfRS) was performed on an orthotopic AY-27 rat bladder urothelial cell carcinoma model to explore potential spectroscopic features revealing neoplastic changes. AY-27 bladder tumor cells were intravesically instilled in four rats and allowed to implant and grow for one week, with two additional rats as the control. A total of 107 SfRS measurements were taken from 27 sites on two control bladders and 80 from four AY-27 treated bladders. The spectral profiles obtained from AY-27 treated bladders revealed various levels of a methemoglobin (MetHb) characteristic spectral feature around 635nm. A multisegment spectral analysis method estimated concentrations of five chromophore compositions including oxyhemoglobin, deoxyhemoglobin, MetHb, lipid and water. The total hemoglobin concentration ([HbT]), the MetHb proportion in the total hemoglobin and the lipid volume content showed possible correlations. The 80 measurements from the AY-27 treated bladders could separate to three sub-sets according to the MetHb proportion. Specifically, 72 were in subset 1 with low proportion (5.3%<[MetHb]<7%), 6 in subset 2 with moderate proportion (7%<[MetHb]<30%), and 2 in subset 3 with significant proportion (>30%). When grouped according to [MetHB], the [HbT] increased from 368 μM of subset 1 to 488 μM of subset 2 to 541 μM of subset 3, in comparison to the 285 μM of the control. The increased total hemoglobin and the elevation of MetHb proportion may signify angiogenesis and degradation in hemoglobin oxygen-transport. Additionally, the lipid volume content decreased from 2.58% in the control to <0.2% in the tumor groups, indicating disruption of subepithelium tissue architecture.

Not only men suffer from sexual dysfunction, but the number of women who have sexual dysfunction rises. Therefore, it is necessary to develop an objective diagnostic technique to examine the sexual dysfunction of female patients, who are afflicted with the disorders. For this purpose, we developed a diffuse optical spectroscopy (DOS) probe to measure the change of oxy-, deoxy-, and total hemoglobin concentration along with blood flow from vaginal wall of female rats. A cylindrical stainless steel DOS probe with a diameter of 3 mm was designed for the vaginal wall of rats which consisted of two lasers (785 and 850nm) and two spectrometers with a separation of 2 mm. A thermistor was placed on the top of the probe to measure the temperature change from vaginal wall during experiments. A modified Beer-Lambert’s law is utilized to acquire the changes of oxy-, deoxy-, and total hemoglobin, and blood flow information is obtained by diffuse speckle contrast analysis technique. For the experiments, Sprague Dawley (~400 g) female rats were divided into two groups (control and vaginal dryness model). Vaginal oxygenation, blood flow and temperature were continuously monitored before and after sexual around induced by apomorphine. After the measurement, histologic examination was performed to support the results from DOS probe in the vaginal wall. The hemodynamic information acquired by the DOS probe can be utilized to establish an objective and accurate standard of the female sexual disorders.

Background: Testicular torsion is an acute urological emergency occurring in children and adolescents. Accurate and fast diagnosis is important as the resulting ischemia can destroy the testis. Currently, Doppler ultrasound is the preferred diagnostic method. Ultrasound is not readily available in all centers which may delay surgical treatment. In this study, a rat model was used to examine the feasibility and sensitivity of using spatially-resolved near infrared spectroscopy (SR-NIRS) with a custom-made miniaturized optical sensor probe to detect and study changes in testicular hemodynamics and oxygenation during three degrees of induced testicular torsion, and after detorsion. Methods: Eight anesthetized rats (16 testes) were studied using SR-NIRS with the miniaturized optical probe applied directly onto the surface of the surgically exposed testis during 360, 720 and 1080 degrees of torsion followed by detorsion. Oxygenated, deoxygenated and total hemoglobin and TOI% were studied pre-and post-manipulations. Results: NIRS monitoring reflected acute testicular ischemia and hypoxia on induction of torsion, and tissue reperfusionreoxygenation after detorsion. Testicular torsion at 720 degrees induced the maximum observed degree of hypoxic changes. In all cases, rhythmic changes were observed in the NIRS signals before inducing torsion; these disappeared after applying 360 degrees of torsion and did not reappear after detorsion. Conclusion: This animal study indicates that SR-NIRS monitoring of the testes using a directly applied miniature sensor is a feasible and sensitive method to detect testicular ischemia and hypoxia immediately after torsion occurs, and testicular reperfusion upon detorsion. This study offers the potential for a SR-NIRS system with a miniaturized sensor to be explored further as a rapid, noninvasive, optical method for detecting testicular torsion in children.

Kidney cancer affects 65,000 new patients every. As computerized tomography became ubiquitous, the number of small, incidentally detected renal masses increased. About 6,000 benign cases are misclassified radiographically as malignant and removed surgically. Raman spectroscopy (RS) has been widely demonstrated for disease discrimination, however intense near-infrared auto-fluorescence of certain tissues (e.g kidney) can present serious challenges to bulk tissue diagnosis. A 1064nm excitation dispersive detection RS system demonstrated the ability to collect spectra with superior quality in tissues with strong auto-fluorescence. Our objective is to develop a 1064 nm dispersive detection RS system capable of differentiating normal and malignant renal tissue. We will report on the design and development of a clinical system for use in nephron sparing surgeries. We will present pilot data that has been collected from normal and malignant ex vivo kidney specimens using a benchtop RS system. A total of 93 measurements were collected from 12 specimens (6 Renal Cell Carcinoma, 6 Normal ). Spectral classification was performed using sparse multinomial logistic regression (SMLR). Correct classification by SMLR was obtained in 78% of the trials with sensitivity and specificity of 82% and 75% respectively. We will present the association of spectral features with biological indicators of healthy and diseased kidney tissue. Our findings indicate that 1064nm RS is a promising technique for differentiation of normal and malignant renal tissue. This indicates the potential for accurately separating healthy and cancerous tissues and suggests implications for utilizing RS for optical biopsy and surgical guidance in nephron sparing surgery.

Limitations of current ureteroscope illumination configurations include presence of shadows and hot spots in images, further degraded by stone debris during laser lithotripsy, which may result in a decrease in stone ablation efficiency, increase in surgical operation time, and potential collateral tissue trauma. Previous studies have reported accidental ureteral tissue perforation from Nitinol stone basket wires during Holmium laser lithotripsy, due in part to poor visibility. Although saline irrigation is routinely used during ureteroscopy to flush stone debris and improve visibility, sub-optimal illumination may still compound these problems. Current illumination geometries and sources are inadequate to produce necessary uniform illumination for optimal visibility and safety during ureteroscopy. By moving away from single and double point source geometry and towards a ring configuration, illumination becomes more uniform in both axes, reducing shadows and increasing depth discernibility. Uric acid and calcium oxalate based stones were chosen for illumination and reflection spectroscopy. Porcine ureters were used as soft tissue samples for comparison. The percent difference in reflection between ureter and stones was greater than 40% for the wavelength ranges of 470-540 nm, and 600-700 nm, making these spectral regions most suitable for high contrast illumination, possibly through narrow band imaging techniques via multiple laser sources and/or optical filters. These improved ureteroscope illumination designs and approaches may potentially reduce complications due to limited visibility during laser lithotripsy and hence increase patient safety.

A miniature ureteroscope has the potential to eliminate need for full anesthesia and dilation, increase comfort and safety of laser lithotripsy via ureteroscopy, and reduce hospital costs via an office based procedure. A prototype, 4.5 Fr (1.5-mm-OD), five channel ureteroscope tip was developed, housing a 200-μm-ID central channel for insertion of small, 100-μm-core fibers and four surrounding channels, each with 510-μm-ID for instrumentation, irrigation, imaging, and illumination, respectively. Common urological instruments (including fibers, guidewires, and stone baskets) were inserted through tip’s working channels to demonstrate feasibility. Low irrigation rates were measured, revealing a need for manual pump-assisted irrigation. Imaging was conducted using 3k, 6k, and 10k pixel miniature flexible endoscopes with 0.4, 0.6, and 0.9 mm outer diameters, respectively. The 3k pixel endoscope with integrated illumination was inserted through the prototype unimpeded, and successfully demonstrated ability to differentiate between hard tissues (e.g. kidney stones) and soft tissues (e.g. ureter wall), for visibility and safety during potential clinical application. Based on both image quality and instrument diameter, the 6k pixel endoscope provided an optimal solution for miniature ureteroscopy.

Native fluorescence spectra are acquired from fresh normal and cancerous human prostate tissues. The fluorescence data are analyzed using a multivariate analysis algorithm such as non-negative matrix factorization. The nonnegative spectral components are retrieved and attributed to the native fluorophores such as collagen, reduced nicotinamide adenine dinucleotide (NADH), and flavin adenine dinucleotide (FAD) in tissue. The retrieved weights of the components, e.g. NADH and FAD are used to estimate the relative concentrations of the native fluorophores and the redox ratio. A machine learning algorithm such as support vector machine (SVM) is used for classification to distinguish normal and cancerous tissue samples based on either the relative concentrations of NADH and FAD or the redox ratio alone. The classification performance is shown based on statistical measures such as sensitivity, specificity, and accuracy, along with the area under receiver operating characteristic (ROC) curve. A cross validation method such as leave-one-out is used to evaluate the predictive performance of the SVM classifier to avoid bias due to overfitting.

Energy-based, radiofrequency and ultrasonic devices provide rapid sealing of blood vessels during laparoscopic procedures. We are exploring infrared lasers as an alternative for vessel sealing with less collateral thermal damage. Previous studies demonstrated vessel sealing in an in vivo porcine model using a 1470-nm laser. However, the initial prototype was designed for open surgery and featured tissue clasping and light delivery mechanisms incompatible with laparoscopic surgery. In this study, a laparoscopic prototype similar to devices in surgical use was developed, and tests were conducted on porcine renal blood vessels. The 5-mm-OD prototype featured a traditional Maryland jaw configuration. Laser energy was delivered through a 550-μm-core fiber and side-delivery from the lower jaw, with beam dimensions of 18-mm-length x 1.2-mm-width. The 1470-nm diode laser delivered 68 W with 3 s activation time. A total of 69 porcine renal vessels with mean diameter of 3.3 ± 1.7 mm were tested, ex vivo. Vessels smaller than 5 mm were consistently sealed (48/51) with burst pressures greater than malignant hypertension blood pressure (180 mmHg), averaging 1038 ± 474 mmHg. Vessels larger than 5 mm were not consistently sealed (6/18), yielding burst pressures of only 174 ± 221 mmHg. Seal width, thermal damage zone, and thermal spread averaged 1.7 ± 0.8, 3.4 ± 0.7, and 1.0 ± 0.4 mm. A novel optical laparoscopic prototype with 5-mm- OD shaft integrated within a standard Maryland jaw design consistently sealed vessels less than 5 mm with minimal thermal spread. Further in vivo studies are planned to test performance across a variety of vessels and tissues.