Tissue Fluorophores Research Articles

Expansion Light Sheet Microscopy Resolves Subcellular Structures in Large Portions of the Songbird Brain.

Expansion microscopy and light sheet imaging (ExLSM) provide a viable alternative to existing tissue clearing and large volume imaging approaches. The analysis of intact volumes of brain tissue presents a distinct challenge in neuroscience. Recent advances in tissue clearing and light sheet microscopy have re-addressed this challenge and blossomed into a plethora of protocols with diverse advantages and disadvantages. While refractive index matching achieves near perfect transparency and allows for imaging at large depths, the resolution of cleared brains is usually limited to the micrometer range. Moreover, the often long and harsh tissue clearing protocols hinder preservation of native fluorescence and antigenicity. Here we image large expanded brain volumes of zebra finch brain tissue in commercially available light sheet microscopes. Our expansion light sheet microscopy (ExLSM) approach presents a viable alternative to many clearing and imaging methods because it improves on tissue processing times, fluorophore compatibility, and image resolution.

Stain-free LED scanning lifetime imaging system for diabetes modified tissue matrices.

In contrast to labor intensive and destructive histological techniques, intrinsic autofluorescence lifetimes of extra cellular matrix proteins can provide label-free imaging of tissue modifications in diseases, including the diabetic ulcers. However, decoupling the complex mixture of tissue fluorophores requires costly and complicated fluorescent lifetime instrumentation. Furthermore, a list of autofluorescent and fluorogenic proteins must be characterized to profile their changes during disease progression. Towards these goals, an imaging system based on frequency domain light-emitting diode (LED) modulation was designed and demonstrated, using off-the-shelf components in a low complexity design. The system was operated by coupling and imaging fluorescence intensities using a pair of objectives. The system's scanning and signal acquisition performances were optimized with respect to etendues. To study fluorescent proteins in diabetic ulcers, lifetimes from purified and pentosidine modified collagen I, collagen III, and elastin were measured. Pentosidine measurements showed a decrease in autofluorescent lifetimes while elevated collagen III in diabetic ulcers showed increased lifetimes. These lifetimes, plus future protein measurements enabled by our system, can serve as standards for developing a biophotonic model of diabetic ulcers. As a proof-of-concept, a 3 cm × 3 cm diabetic foot ulcer was imaged using the developed system. Phasor analysis was applied to aid the interpretation of lifetime images. As a result, a compact biophotonic imaging system targeting diabetic tissue was achieved, towards making the technique accessible for clinical histology.

Noninvasive Optical Assessment of Implanted Engineered Tissues Correlates with Cytokine Secretion.

Fluorescence lifetime sensing has been shown to noninvasively characterize the preimplantation health and viability of engineered tissue constructs. However, current practices to monitor postimplantation construct integration are either qualitative (visual assessment) or destructive (tissue histology). We employed label-free fluorescence lifetime spectroscopy for quantitative, noninvasive optical assessment of engineered tissue constructs that were implanted into a murine model. The portable system was designed to be suitable for intravital measurements and included a handheld probe to precisely and rapidly acquire data at multiple sites per construct. Our model tissue constructs were manufactured from primary human cells to simulate patient variability based on a standard protocol, and half of the manufactured constructs were stressed to create a range of health states. Secreted amounts of three cytokines that relate to cellular viability were measured in vitro to assess preimplantation construct health: interleukin-8 (IL-8), human β-defensin 1 (hBD-1), and vascular endothelial growth factor (VEGF). Preimplantation cytokine secretion ranged from 1.5 to 33.5 pg/mL for IL-8, from 3.4 to 195.0 pg/mL for hBD-1, and from 0.1 to 154.3 pg/mL for VEGF. In vivo optical sensing assessed constructs at 1 and 3 weeks postimplantation. We found that at 1 week postimplantation, in vivo optical parameters correlated with in vitro preimplantation secretion levels of all three cytokines (p < 0.05). This correlation was not observed in optical measurements at 3 weeks postimplantation when histology showed that the constructs had re-epithelialized, independent of preimplantation health state, supporting the lack of a correlation. These results suggest that clinical optical diagnostic tools based on label-free fluorescence lifetime sensing of endogenous tissue fluorophores could noninvasively monitor postimplantation integration of engineered tissues.

QF-SSOP: real-time optical property corrected fluorescence imaging.

Fluorescence imaging is well suited to provide image guidance during resections in oncologic and vascular surgery. However, the distorting effects of tissue optical properties on the emitted fluorescence are poorly compensated for on even the most advanced fluorescence image guidance systems, leading to subjective and inaccurate estimates of tissue fluorophore concentrations. Here we present a novel fluorescence imaging technique that performs real-time (i.e., video rate) optical property corrected fluorescence imaging. We perform full field of view simultaneous imaging of tissue optical properties using Single Snapshot of Optical Properties (SSOP) and fluorescence detection. The estimated optical properties are used to correct the emitted fluorescence with a quantitative fluorescence model to provide quantitative fluorescence-Single Snapshot of Optical Properties (qF-SSOP) images with less than 5% error. The technique is rigorous, fast, and quantitative, enabling ease of integration into the surgical workflow with the potential to improve molecular guidance intraoperatively.

A fast forward solver of fluorescence diffuse optical tomography based on the lattice Boltzmann method.

Fluorescence diffuse optical tomography (FDOT) is a new molecular imaging technology, which uses near-infrared light to excite the fluorophore in tissues. According to the measurements detected on the surface of imaged object, the fluorescent quantum yield as well as lifetime of the fluorescence can be reconstructed. However, the reconstruction of FDOT remains a challenging problem because the conventional forward solvers are time consuming. Thus, a forward model solver that would enable the fast imaging is necessary. This paper describes a new forward solver to simulate the propagation of photons in tissues based on the lattice Boltzmann method (LBM). This is accomplished by propagation photons in tissues guided by the LBM. To evaluate the performance of the proposed LBM, based on the numerical simulation, we compared the light distribution generated by the LBM with the diffusion equation implemented by COMSOL in four different cases. The experimental results indicate that compared to diffusion equation, the LBM can reduce the computation time for the forward solver of FDOT while preserving the similar accuracy.

Development of thin skin mimicking bilayer solid tissue phantoms for optical spectroscopic studies.

In vivo spectroscopic measurements have the proven potential to provide important insight about the changes in tissue during the development of malignancies and thus help to diagnose tissue pathologies. Extraction of intrinsic data in the presence of varying amounts of scatterers and absorbers offers great challenges in the development of such techniques to the clinical level. Fabrication of optical phantoms, tailored to the biochemical as well as morphological features of the target tissue, can help to generate a spectral database for a given optical spectral measurement system. Such databases, along with appropriate pattern matching algorithms, could be integrated with in vivo measurements for any desired quantitative analysis of the target tissue. This paper addresses the fabrication of such soft, photo stable, thin bilayer phantoms, mimicking skin tissue in layer dimensions and optical properties. The performance evaluation of the fabricated set of phantoms is carried out using a portable fluorescence spectral measurement system. The alterations in flavin adenine dinucleotide (FAD)-a tissue fluorophore that provides important information about dysplastic progressions in tissues associated with cancer development based on changes in emission spectra-fluorescence with varied concentrations of absorbers and scatterers present in the phantom are analyzed and the results are presented. Alterations in the emission intensity, shift in emission wavelength and broadening of the emission spectrum were found to be potential markers in the assessment of biochemical changes that occur during the progression of dysplasia.

Predictive assessment of kidney functional recovery following ischemic injury using optical spectroscopy.

Functional changes in rat kidneys during the induced ischemic injury and recovery phases were explored using multimodal autofluorescence and light scattering imaging. The aim is to evaluate the use of noncontact optical signatures for rapid assessment of tissue function and viability. Specifically, autofluorescence images were acquired in vivo under 355, 325, and 266 nm illumination while light scattering images were collected at the excitation wavelengths as well as using relatively narrowband light centered at 500 nm. The images were simultaneously recorded using a multimodal optical imaging system. The signals were analyzed to obtain time constants, which were correlated to kidney dysfunction as determined by a subsequent survival study and histopathological analysis. Analysis of both the light scattering and autofluorescence images suggests that changes in tissue microstructure, fluorophore emission, and blood absorption spectral characteristics, coupled with vascular response, contribute to the behavior of the observed signal, which may be used to obtain tissue functional information and offer the ability to predict posttransplant kidney function.

Jones-matrix mapping of optically anisotropic fluorophores of molecular biological tissues in the diagnostics of death causes

Jones-matrix mapping of optically anisotropic fluorophores of molecular biological tissues in the diagnostics of death causes

A Dual-Modality System for Both Multi-Color Ultrasound-Switchable Fluorescence and Ultrasound Imaging.

Simultaneous imaging of multiple targets (SIMT) in opaque biological tissues is an important goal for molecular imaging in the future. Multi-color fluorescence imaging in deep tissues is a promising technology to reach this goal. In this work, we developed a dual-modality imaging system by combining our recently developed ultrasound-switchable fluorescence (USF) imaging technology with the conventional ultrasound (US) B-mode imaging. This dual-modality system can simultaneously image tissue acoustic structure information and multi-color fluorophores in centimeter-deep tissue with comparable spatial resolutions. To conduct USF imaging on the same plane (i.e., x-z plane) as US imaging, we adopted two 90°-crossed ultrasound transducers with an overlapped focal region, while the US transducer (the third one) was positioned at the center of these two USF transducers. Thus, the axial resolution of USF is close to the lateral resolution, which allows a point-by-point USF scanning on the same plane as the US imaging. Both multi-color USF and ultrasound imaging of a tissue phantom were demonstrated.

Depth-dependent autofluorescence photobleaching using 325, 473, 633, and 785nm of porcine ear skin ex vivo.

Autofluorescence photobleaching describes the decrease of fluorescence intensity of endogenous fluorophores in biological tissue upon light irradiation. The origin of autofluorescence photobleaching is not fully understood. In the skin, the spatial distribution of various endogenous fluorophores varies within the skin layers. Most endogenous fluorophores are excited in the ultraviolet and short visible wavelength range, and only a few, such as porphyrins (red) and melanin (near-infrared), are excited at longer wavelengths. The excitation wavelength- and depth-dependent irradiation of skin will therefore excite different fluorophores, which will likely influence the photobleaching characteristics. The autofluorescence photobleaching of porcine ear skin has been measured ex vivo using 325, 473, 633, and 785 nm excitation at different skin depths from the surface to the dermis at 150 ? ? m . Confocal Raman microscopes were used to achieve sufficient spatial resolution of the measurements. The autofluorescence area under the curve was measured for 21 consecutive acquisitions of 15 s. In all cases, the photobleaching follows a two-exponential decay function approximated by nonlinear regression. The results show that photobleaching can be applied to improve the signal-to-noise ratio in Raman spectroscopy for all of the applied excitation wavelengths and skin depths.

Excitation – emission matrices and synchronous fluorescence spectroscopy for cancer diagnostics in gastrointestinal tract

We report the development of an improved fluorescence technique for cancer diagnostics in the gastrointestinal tract. We investigate the fluorescence of ex vivo colorectal (cancerous and healthy) tissue samples using excitation – emission matrix (EEM) and synchronous fluorescence spectroscopy (SFS) steady-state approaches. The obtained results are processed for revealing characteristic fluorescence spectral features with a valuable diagnostic meaning. The main tissue fluorophores, contributing to the observed fluorescence, are tyrosine, tryptophan, NADH, FAD, collagen and elastin. Based on the results of the Mann – Whitney test as useful parameters for differentiation of gastrointestinal cancer from normal mucosa, we suggest using excitation wavelengths in the range for fluorescence spectroscopy and wavelengths intervals of and for SFS.

Wireless fluorescence capsule for endoscopy using single photon-based detection.

Fluorescence Imaging (FI) is a powerful technique in biological science and clinical medicine. Current FI devices that are used either for in-vivo or in-vitro studies are expensive, bulky and consume substantial power, confining the technique to laboratories and hospital examination rooms. Here we present a miniaturised wireless fluorescence endoscope capsule with low power consumption that will pave the way for future FI systems and applications. With enhanced sensitivity compared to existing technology we have demonstrated that the capsule can be successfully used to image tissue autofluorescence and targeted fluorescence via fluorophore labelling of tissues. The capsule incorporates a state-of-the-art complementary metal oxide semiconductor single photon avalanche detector imaging array, miniaturised optical isolation, wireless technology and low power design. When in use the capsule consumes only 30.9 mW, and deploys very low-level 468 nm illumination. The device has the potential to replace highly power-hungry intrusive optical fibre based endoscopes and to extend the range of clinical examination below the duodenum. To demonstrate the performance of our capsule, we imaged fluorescence phantoms incorporating principal tissue fluorophores (flavins) and absorbers (haemoglobin). We also demonstrated the utility of marker identification by imaging a 20 μM fluorescein isothiocyanate (FITC) labelling solution on mammalian tissue.

Intravital autofluorescence 2-photon microscopy of murine intestinal mucosa with ultra-broadband femtosecond laser pulse excitation: image quality, photodamage, and inflammation.

Ultra-broadband excitation with ultrashort pulses may enable simultaneous excitation of multiple endogenous fluorophores in vital tissue. Imaging living gut mucosa by autofluorescence 2-photon microscopy with more than 150 nm broad excitation at an 800-nm central wavelength from a sub-10 fs titanium-sapphire (Ti:sapphire) laser with a dielectric mirror based prechirp was compared to the excitation with 220 fs pulses of a tunable Ti:sapphire laser at 730 and 800 nm wavelengths. Excitation efficiency, image quality, and photochemical damage were evaluated. At similar excitation fluxes, the same image brightness was achieved with both lasers. As expected, with ultra-broadband pulses, fluorescence from NAD(P)H, flavines, and lipoproteins was observed simultaneously. However, nonlinear photodamage apparent as hyperfluorescence with functional and structural alterations of the tissue occurred earlier when the laser power was adjusted to the same image brightness. After only a few minutes, the immigration of polymorphonuclear leucocytes into the epithelium and degranulation of these cells, a sign of inflammation, was observed. Photodamage is promoted by the higher peak irradiances and/or by nonoptimal excitation of autofluorescence at the longer wavelength. We conclude that excitation with a tunable narrow bandwidth laser is preferable to ultra-broadband excitation for autofluorescence-based 2-photon microscopy, unless the spectral phase can be controlled to optimize excitation conditions.

Meshless reconstruction method for fluorescence molecular tomography based on compactly supported radial basis function.

Fluorescence molecular tomography (FMT) is a promising tool in the study of cancer, drug discovery, and disease diagnosis, enabling noninvasive and quantitative imaging of the biodistribution of fluorophores in deep tissues via image reconstruction techniques. Conventional reconstruction methods based on the finite-element method (FEM) have achieved acceptable stability and efficiency. However, some inherent shortcomings in FEM meshes, such as time consumption in mesh generation and a large discretization error, limit further biomedical application. In this paper, we propose a meshless method for reconstruction of FMT (MM-FMT) using compactly supported radial basis functions (CSRBFs). With CSRBFs, the image domain can be accurately expressed by continuous CSRBFs, avoiding the discretization error to a certain degree. After direct collocation with CSRBFs, the conventional optimization techniques, including Tikhonov, L1-norm iteration shrinkage (L1-IS), and sparsity adaptive matching pursuit, were adopted to solve the meshless reconstruction. To evaluate the performance of the proposed MM-FMT, we performed numerical heterogeneous mouse experiments and in vivo bead-implanted mouse experiments. The results suggest that the proposed MM-FMT method can reduce the position error of the reconstruction result to smaller than 0.4 mm for the double-source case, which is a significant improvement for FMT.

Human feasibility study of fluorescence spectroscopy guided optical biopsy needle for prostate cancer diagnosis.

Current prostate biopsy cores have a very low diagnostic yield. These biopsies often fail to diagnose prostate cancer since 90% of cores are histopathologically classified as benign. The concentrations of endogenous fluorophores in prostate tissue vary with disease states. Thus, fluorescence spectroscopy could be utilized to quantify these variations for identification of malignant lesions. We investigated clinical feasibility of a 14 gauge (1.98 mm) optical biopsy needle guided by fluorescence spectroscopy for real-time in vivo prostate cancer diagnosis. Built-in optical sensor has 8×100μm fibers for tissue excitation and a single 200μm fiber to collect spectral data. Custom-made fluorometer has 2 light-emitting diodes at 290 and 340 nm and a spectrometer. User interface for fluorometer operation and data collection was developed using LabView software. Each spectral data acquisition required ~2 seconds. The in vivo biopsies were performed during radical retropubic prostatectomy surgery on the exposed prostate with blood flow to the gland intact. A tissue biopsy core was obtained from each biopsy site after acquisition of spectral data. Above procedure was repeated ex vivo after surgical excision of the prostate. Biopsy cores were histopathologically classified as either benign or malignant and correlated with corresponding spectral data. Partial Least Square analysis was performed to determine diagnostically significant principal components as potential classifiers. A linear support vector machine and leave-one-out cross validation method was employed for tissue classification. Thirteen patients were consented to the study. Histopathological analysis found cancer in 29/208 in vivo and 51/224 ex vivo viable biopsy cores. Study results show 72% sensitivity, 66% specificity, and 93% negative predictive value for in vivo and 75%, 80%, and 93%, respectively, for ex vivo malignant versus benign prostatic tissue classification. Optical biopsy needle has a very high negative predictive value to indicate benign tissue while sufficient sensitivity for targeting areas suspicious for cancer within the prostate gland. Hence, the optical biopsy needle can increase the diagnostic yield of prostate biopsies with consequent improvement in patient care.

Nonlinear Behavior of the Autofluorescence Intensity on the Surface of Light-Scattering Biotissues and its Theoretical Proof.

In the up-to-date medical laser fluorescence spectroscopy (LFS) in vivo, there is a problem of quantification of fluorophores concentrations in optically-turbid biotissues by measurements of the laser induced autofluorescence flux on the surface of these tissues. One of the main problems is: whether the flux depends linearly or non-linearly on the concentration of fluorophores in tissues? The purpose of this work was both experimental and theoretical study of the character of dependencies between measured fluorescence intensities and fluorophores concentrations in optically-turbid media. In the experimental part of our study, measurements of the superficial fluorescence on phantoms at various known concentrations of fluorophores in them were carried out. As a result, experimental dependencies of registered intensities of the laser-induced autofluorescence emission were plotted against fluorophore concentrations. In the theoretical part of the study, the analytical solution for the fluorescence emission by Kokhanovsky's method based on the well-known two-flux Kubelka-Munk approach (KMA) was used. In addition, in our study the Kokhanovsky's method was modified by its association with our improved KMA, allowing us to receive exact analytical solutions for boundary intensities collected by optical probes. As a result, a set of theoretical curves describing the influence of fluorophore concentrations on the registered autofluorescence intensities was obtained, as well. Both experimental and theoretical results show a good qualitative agreement with each other. Also, these results demonstrate that the dependence of the fluorescence intensity on tissues' optical properties and on the concentration of fluorophores in light-scattering tissues can be both nonlinear and non-monotonic.

MP48-02 FLUORESCENCE SPECTROSCOPY CAN INCREASE DIAGNOSTIC YIELD OF PROSTATE BIOPSIES

You have accessJournal of UrologyProstate Cancer: Detection and Screening I1 Apr 2015MP48-02 FLUORESCENCE SPECTROSCOPY CAN INCREASE DIAGNOSTIC YIELD OF PROSTATE BIOPSIES E. David Crawford, Edward A. Jasion, Yongjun Liu, John Daily, Paul Arangua, Clifford Jones, S. Russell Nash, and Priya Werahera E. David CrawfordE. David Crawford More articles by this author , Edward A. JasionEdward A. Jasion More articles by this author , Yongjun LiuYongjun Liu More articles by this author , John DailyJohn Daily More articles by this author , Paul AranguaPaul Arangua More articles by this author , Clifford JonesClifford Jones More articles by this author , S. Russell NashS. Russell Nash More articles by this author , and Priya WeraheraPriya Werahera More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.1676AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Transrectal ultrasound (TRUS) guided prostate biopsy cores have a very low diagnostic yield as 90% cores are histopathologically classified as benign. We investigated potential clinical application fluorescence spectroscopy (FS) to increase diagnostic yield of TRUS biopsies. The fluorescence emissions from natural fluorophores (e.g., collagen) in prostate tissue are altered by the presence of prostate cancer (PCa). Thus, FS can be used to distinct malignant locations from benign to aid TRUS biopsies. METHODS 14G optical biopsy needle was prototyped to capture FS of prostate tissue. Integrated optical sensor uses 8x100 μm fibers for excitation and 1x200 μm fiber to collect FS data. Custom made fluorometers with 2 light-emitting diodes at 290 and 340 nm and a spectrometer were used to measure FS of prostate tissue. User interface for fluorometer operation and data collection was developed using LabView software. Each spectral data acquisition takes ∼250 milliseconds. The in vivo biopsies were performed during radical prostatectomy surgery on the exposed prostate with blood flow to the gland intact. A tissue biopsy core was obtained from each biopsy site after acquisition of FS data. Above procedure was repeated ex vivo after surgical excision of the prostate. Biopsy cores were histopathologically classified as benign or malignant and correlated with corresponding FS data. Partial Least Square analysis of the FS data was performed to determine principal components (PCs) at each excitation wavelength. Using linear support vector machine and leave-one-out cross validation method, several combinations of PCs were tested for their ability to classify benign vs. malignant prostatic tissue. RESULTS Thirteen patients were consented. A total of 208 in vivo biopsies (29 malignant) and 224 ex vivo biopsies (51 malignant) were analyzed. Results for benign vs. malignant prostatic tissue classification based on number of PCs used are given in the table. CONCLUSIONS Our optical biopsy needle assisted with FS has a very high NPV to indicate benign tissue while sufficient SE and SP for real-time diagnosis of PCa by targeting areas positive for cancer within the prostate gland. Hence, systematic use of optical biopsies can potentially increase diagnostic yield of prostate biopsies. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e594 Peer Review Report Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information E. David Crawford More articles by this author Edward A. Jasion More articles by this author Yongjun Liu More articles by this author John Daily More articles by this author Paul Arangua More articles by this author Clifford Jones More articles by this author S. Russell Nash More articles by this author Priya Werahera More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

O01 * A META-ANALYSIS OF THE DIAGNOSTIC ABILITY, EFFICACY, SAFETY AND COST EFFECTIVENESS OF 5-AMINOLEVULINIC ACID GUIDED RESECTION OF HIGH GRADE GLIOMAS.

INTRODUCTION: It is well appreciated that maximal resection of glioblastomas improves outcome but is difficult to achieve due to the invasive nature of gliomas. 5-aminolevulinic acid (5-ALA) improves tumour detection by selective conversion to a fluorophore in tumour tissue that is detected by fluorescence under blue light. This meta-analysis aims to assess the ability of 5-ALA as a tool for identifying tumour as well as the efficacy, safety and cost-effectiveness of 5-ALA resections. METHOD: A systematic review of PubMed and EMBASE was was supplemented with review of the trials registry and expert opinion. Case reports and reviews were excluded. Studies reporting diagnostic accuracy, clinical efficacy (extent of resection or progression free survival [PFS]), safety or cost effectiveness in adults with resected high grade gliomas were included. RESULTS: 52 studies were identified of which 21 fulfilled the search criteria. The diagnostic accuracy of fluorescence identifying tumour had 87% sensitivity, 93% specificity, 98% positive predictive value and 58% negative predictive value. 72.3% of patients had a complete resection of the enhancing tumour. Two studies demonstrated a prolonged PFS compared to a white light group. There were no increases in morbidity in patients without deficits pre-operatively. 5-ALA costs £1,241.56 more per patient and this is largely due to patients receiving more cycles of chemotherapy. The cost per QALY was £7,424.46 - well below the NICE threshold. CONCLUSION: This meta-analysis shows that 5-ALA is an accurate diagnostic tool, improves progression free survival but does not increase morbidity and is cost effective. This should now be considered as a standard of care.

Mueller-matrix mapping of optically anisotropic fluorophores of biological tissues in the diagnosis of cancer

We report the results of studying the polarisation manifestations of laser autofluorescence of optically anisotropic structures in biological tissues. A Mueller-matrix model is proposed to describe their complex anisotropy (linear and circular birefringence, linear and circular dichroism). The relationship is established between the mechanisms of optical anisotropy and polarisation manifestations of laser autofluorescence of histological sections of rectal tissue biopsy in different spectral regions. The ranges of changes in the statistical moments of the 1st-to-4th orders, which describe the distribution of the azimuth-invariant elements of Mueller matrices of rectal tissue autofluorescence, are found. Effectiveness of laser autofluorescence polarimetry is determined and the histological sections of biopsy of benign (polyp) and malignant (adenocarcinoma) tumours of the rectal wall are differentiated for the first time.

Systematic diagnosis of prostate cancer using an optical biopsy needle adjunct with fluorescence spectroscopy.

Transrectal ultrasound guided prostate biopsies often fail to diagnose prostate cancer with 90% of cores reported as benign. Thus, it is desirable to target prostate cancer lesions while reducing the sampling of benign tissue. The concentrations of natural fluorophores in prostate tissue fluctuate with disease states. Hence, fluorescence spectroscopy could be used to quantify these fluctuations to identify prostate cancer. An optical biopsy needle with a light sensitive optical probe at the tip of the inner needle was developed to take prostate biopsies after measuring tissue fluorescence with a laboratory fluorometer. The optical probe consists of eight 100 μm fibers for tissue excitation and a single 200 μm fiber to capture fluorescence spectra. Random biopsy cores were taken from 20 surgically excised prostates after measuring fluorescence spectra of tissue between 295-550nm for several excitations between 280-350nm. Each biopsy core was histopathologically classified and correlated with corresponding spectra. Prostate biopsies were grouped into benign or malignant based on the histological findings. Out of 187 biopsy cores, 109 were benign and 78 were malignant. Partial least square analysis of tissue spectra was performed to identify diagnostically significant principal components as potential classifiers. A linear support vector machine and leave-one-out cross validation method was employed for tissue classification. Study results show 86% sensitivity, 87% specificity, 90% negative predictive value, and 83% positive predictive value for benign versus malignant prostate tissue classification. This study demonstrates potential clinical applications of fluorescence spectroscopy guided optical biopsy needle for prostate cancer diagnosis with the consequent improvement of patient care.