This PDF file contains the front matter associated with SPIE Proceedings Volume 9698, including the Title Page, Copyright information, Table of Contents, Introduction, and the Conference Committee listing.

Searching for new methods to provide information of biochemical composition and structure is critical to improve the prognosis of thyroid diseases. The use of time-resolved fluorescence techniques to detect biochemical composition and tissue structure alterations could help develop a portable, minimally invasive, and non-destructive method to assist during surgical procedures. This research looks for employ a fluorescence technique based on lifetime measurements to differentiate healthy and benign lesions from malignant thyroid tissue. We employ a wide range of excitation and chose a more appropriate region for this work: 298-300 nm; and the fluorescence decay was measured at 340-450 nm. We observed fluorescence lifetimes at 340 nm emission of 0.80±0.26 and 3.94±0.47 ns for healthy tissue; 0.90±0.24 and 4.05±0.46 ns for benign lesions; and 1.21±0.14 and 4.63±0.25 ns for malignant lesions. For 450 nm emissions, we obtain lifetimes of 0.25±0.18 and 3.99±0.39 ns for healthy tissue, 0.24±0.17 and 4.20±0.48 ns for benign lesions, 0.33±0.32 and 4.55±0.55 ns for malignant lesions. We successfully demonstrated that fluorescence lifetimes at 340 nm emission can differentiate between thyroid malignant and healthy/benign tissues.

Barrett’s esophagus is a condition that predisposes patients to esophageal cancer. Early detection of cancer in these patients can be curative, but is confounded by a lack of contrast in white light endoscopy (WLE). Application of fluorescently-labeled lectins to the esophagus during endoscopy can more accurately delineate dysplasia emerging within Barrett’s than WLE1, but strong tissue autofluorescence has limited sensitivity and dynamic range of this approach. To overcome this challenge, we synthesized a near-infrared (NIR) fluorescent lectin and have constructed a clinically translatable endoscope for simultaneous WLE and NIR imaging. An imaging fiber bundle, shielded from patient contact using a disposable catheter, relays collected light into an optical path that splits the WL reflectance and NIR emission onto two cameras for simultaneous video-rate recording. The captured images are co-registered and the honeycomb artifact arising from the fiber bundle is removed using interpolation between image points derived from individual fibers. A minimum detectable concentration of 110 nM was determined using a dilution series of IRDye800CW-lectin in black well plates. We have demonstrated the ability to use our endoscope to distinguish between different tissue types in ex vivo mouse stomachs. Future work using human ex vivo tissue specimens will determine safe illumination limits and sensitivity for dysplasia and adenocarcinoma in Barrett’s esophagus, prior to commencing clinical trials.

Endogenous fluorescence lifetime imaging (FLIM) provides direct access to the concomitant functional and biochemical changes accompanying tissue transition from benign to precancerous and cancerous. Since FLIM can noninvasively measure different and complementary biomarkers of precancer and cancer, we hypothesize that it will aid in clinically detecting early oral epithelial cancer. Our group has recently demonstrated the detection of benign from premalignant and malignant lesions based on endogenous multispectral FLIM in the hamster cheek-pouch model. Encouraged by these positive preliminary results, we have developed a handheld endoscope capable of acquiring multispectral FLIM images in real time from the oral mucosa. This novel FLIM endoscope is being used for imaging clinically suspicious pre-malignant and malignant lesions from patients before undergoing tissue biopsy for histopathological diagnosis of oral epithelial cancer. Our preliminary results thus far are already suggesting the potential of endogenous FLIM for distinguishing a variety of benign lesions from advanced dysplasia and squamous cell carcinoma (SCC). To the best of out knowledge, this is the first in vivo human study aiming to demonstrate the ability to predict the true malignancy of clinically suspicious lesions using endogenous FLIM. If successful, the resulting clinical tool will allow noninvasive real-time detection of epithelial precancerous and cancerous lesions in the oral mucosa and could potentially be used to assist at every step involved on the clinical management of oral cancer patients, from early screening and diagnosis, to treatment and monitoring of recurrence.

Raman spectroscopy has been proven to have tremendous potential as biomedical analytical tool for spectroscopic disease diagnostics. The use of fiberoptic coupled Raman spectroscopy systems can enable in-vivo characterization of suspicious lesions. However, Raman spectroscopy has the drawback of rather long acquisition times of several hundreds of milliseconds which makes scanning of larger regions quite challenging. By combining Raman spectroscopy with a fast imaging technique this problem can be alleviate in part. Fluorescence lifetime imaging (FLIm) offers a great potential for such a combination. FLIm can allow for fast tissue area pre-segmentation and location of the points for Raman spectra acquisition. Here, we introduce an optical fiber probe combining FLIm and Raman spectroscopy with an outer diameter of 2 mm. Fluorescence is generated via excitation with a fiber laser at 355 nm. The fluorescence emission is spectrally resolved using a custom-made wavelength-selection module (WSM). The Raman excitation power at 785 nm was set to 50 mW for the in-vivo measurements to prevent sample drying. The lateral probe resolution was determined to be <250 μm for both modalities. This value was taken as step size for several raster scans of different tissue types which were conducted to show the overlap of both modalities under realistic conditions. Finally the probe was used for in vivo raster scans of a rat’s brain and subsequently to acquire FLIm guided Raman spectra of several tissues in and around the craniotomy.

In the clinical daily life various lesions of the oral cavity have shown different aspects, generating an inconclusive or doubtful diagnosis. In general, oral injuries are diagnosed by histopathological analysis from biopsy, which is an invasive procedure and does not gives immediate results. In the other hand, Raman spectroscopy technique it is a real time and minimal invasive analytical tool, with notable diagnostic capability. This study aims to characterize, by optical fiber Raman-based spectroscopy (OFRS), normal, inflammatory, potentially malignant, benign and malign oral lesions. Raman data were collected by a Holospec f / 1.8 spectrograph (Kayser Optical Systems) coupled to an optical fiber, with a 785nm laser line source and a CCD Detector. The data were pre-processed and vector normalized. The average analysis and standard deviation was performed associated with cluster analysis and compared to the histopalogical results. Samples of described oral pathological processes were used in the study. The OFRS was efficient to characterized oral lesions and normal mucosa, in which biochemical information related to vibrational modes of proteins, lipids, nucleic acids and carbohydrates were observed. The technique (OFRS) is able to demonstrate biochemical information concern different types of oral lesions showing that Raman spectroscopy could be useful for an early and minimal invasive diagnosis.

Radiation therapy is a principal modality for head and neck cancers and its efficacy depends on tumor hemodynamics. Our laboratory developed a hybrid diffuse optical instrument allowing for simultaneous measurements of tumor blood flow and oxygenation. In this study, the clinically involved cervical lymph node was monitored by the hybrid instrument once a week over the treatment period of seven weeks. Based on treatment outcomes within one year, patients were classified into a complete response group (CR) and an incomplete response group (IR) with remote metastasis and/or local recurrence. A linear mixed models was used to compare tumor hemodynamic responses to the treatment between the two groups. Interestingly, we found that human papilloma virus (HPV-16) status largely affected tumor hemodynamic responses. For HPV-16 negative tumors, significant differences in blood flow index (BFI, p = 0.007) and reduced scattering coefficient (μs’, p = 0.0005) were observed between the two groups; IR tumors exhibited higher μs’ values and a continuous increase in BFI over the treatment period. For HPV-16 positive tumors, oxygenated hemoglobin concentration ([HbO2]) and blood oxygen saturation (StO2) were significant different (p = 0.003 and 0.01, respectively); IR group showed lower [HbO2] and StO2. Our results imply HPV-16 negative tumors with higher density of vasculature (μs’) and higher blood flow show poor responses to radiotherapy and HPV-16 positive tumors with lower tissue oxygenation level (lower StO2 and [HbO2]) exhibit poor treatment outcomes. Our diffuse optical measurements show the great potential for early prediction of radiotherapy in head and neck cancers.

We report here on the ability of elastic light scattering in discriminating Gram+, Gram- and yeasts at an early stage of growth (6h). Our technique is non-invasive, low cost and does require neither skilled operators nor reagents. Therefore it is compatible with automation. It is based on the analysis of the scattering pattern (scatterogram) generated by a bacterial microcolony growing on agar, when placed in the path of a laser beam. Measurements are directly performed on closed Petri dishes.

The characteristic features of a given scatterogram are first computed by projecting the pattern onto the Zernike orthogonal basis. Then the obtained data are compared to a database so that machine learning can yield identification result. A 10-fold cross-validation was performed on a database over 8 species (15 strains, 1906 scatterograms), at 6h of incubation. It yielded a 94% correct classification rate between Gram+, Gram- and yeasts. Results can be improved by using a more relevant function basis for projections, such as Fourier-Bessel functions. A fully integrated instrument has been installed at the Grenoble hospital’s laboratory of bacteriology and a validation campaign has been started for the early screening of MSSA and MRSA (Staphylococcus aureus, methicillin-resistant S. aureus) carriers.

Up to now, all the published studies about elastic scattering were performed in a forward mode, which is restricted to transparent media. However, in clinical diagnostics, most of media are opaque, such as blood-supplemented agar. That is why we propose a novel scheme capable of collecting back-scattered light which provides comparable results.

Extracting biochemical information from tissue autofluorescence is a promising approach to non-invasively monitor disease treatments at a cellular level, without using any external biomarkers. Our recently developed unsupervised hyperspectral unmixing by Dependent Component Analysis (DECA) provides robust and detailed metabolic information with proper account of intrinsic cellular heterogeneity. Moreover this method is compatible with established methods of fluorescent biomarker labelling. Recently adipose-derived stem cell (ADSC) – based therapies have been introduced for treating different diseases in animals and humans. ADSC have been shown promise in regenerative treatments for osteoarthritis and other bone and joint disorders. One of the mechanism of their action is their anti-inflammatory effects within osteoarthritic joints which aid the regeneration of cartilage. These therapeutic effects are known to be driven by secretions of different cytokines from the ADSCs. We have been using the hyperspectral unmixing techniques to study in-vitro the effects of ADSC-derived cytokine-rich secretions with the cartilage chip in both human and bovine samples. The study of metabolic effects of different cytokine treatment on different cartilage layers makes it possible to compare the merits of those treatments for repairing cartilage.

This study was to evaluate the feasibility of near infrared (NIR) fluorescent images as a tool for evaluating the perfusion of the gastric tube after esophagectomy. In addition, we investigated the time required to acquire enough signal to confirm the presence of ischemia in gastric tube after injection of indocyanine green (ICG) through peripheral versus and central venous route. 4 porcine underwent esophagogastrostomy and their right gastric arteries were ligated to mimic ischemic condition of gastric tube. ICG (0.6mg/kg) was intravenously injected and the fluorescence signal-to-background ratios (SBR) were measured by using the custom-built intraoperative color and fluorescence imaging system (ICFIS). We evaluated perfusion of gastric tubes by comparing their SBR with esophageal SBR. In ischemic models, SBR of esophagus was higher than that of gastric tube (2.8±0.54 vs. 1.7±0.37, p<0.05). It showed high esophagus-stomach signal to signal ratio. (SSR, 1.8±0.76). We also could observe recovery of blood perfusion in few minutes after releasing the ligation of right gastric artery. In addition, in comparison study according to the injection route of ICG, The time to acquire signal stabilization was faster in central than in peripheral route (119 ± 65.1 seconds in central route vs. 295±130.4 in peripheral route, p<0.05). NIR fluorescent images could provide the real-time information if there was ischemia or not in gastric tube during operation. And, central injection of ICG might give that information faster than peripheral route.

Currently available pathology techniques for obtaining a rapid tissue diagnosis, or for determining the adequacy of specimens intended for downstream analysis, are too slow, labor-intensive, and destructive for point-of-care (POC) applications. We previously demonstrated video-rate structured illumination microscopy (VR-SIM) for accurate, high-throughput, non-destructive diagnostic imaging of fluorescently-stained prostate biopsies in seconds per biopsy, with an area under the ROC curve of 0.82-0.88 after pathologist review. In addition, we have demonstrated that it is feasible to use VR-SIM to routinely image very large gross pathology specimens, such as entire prostate resection surfaces, in relatively short timeframes at subcellular resolution. However, our prior work has focused on applications in prostate cancer; the utility in other organ sites has not been explored. Here we extended our technology to varying size kidney, liver, and lung biopsies. We conducted a validation study of VR-SIM against histopathology on a variety of human tissues, including both small biopsies and large slices of tissue. We conducted a blinded study in which the study pathologist accurately identified the organs based on VR-SIM images alone. The results were then used to create a clinical atlas between VR-SIM and H and E images for the different tissues of interest. This clinical atlas will be used to aid in pathologist interpretation in future POC clinical applications of VR-SIM in kidney, liver, and lung. Such applications could include on-site identification of the presence of kidney glomeruli for to ensure successful downstream IHC analysis, or determination of the adequacy of lung cancer biopsies for genomic analysis.

Cervical cancer is still one of the most relevant women cancer types, since the 5-year survival rate is of only around 68%. Prevention and early diagnosis are the best strategies to improve cervical cancer prognosis. Conventional diagnosis procedure in Gynecology is mainly based on the macroscopic clinical evaluation, Pap smear cytology, and biopsy, if needed. A portable microscope with dual configuration and its use for diagnosis in Gynecology is investigated. The microscope has interchangeable parts that allow its use for cytopathology smear samples or in situ endoscopic tissue interrogation, both using acriflavine as a nuclei marker. Patients of the Women Ambulatory of the School of Medicine (UNIARA, Araraquara, Brazil) were interrogated during the colposcopy examination. The cervix was initially cleaned using an acetic acid solution, and a 0.05% (wt/vol) acriflavine in saline solution was topically applied at the tissue surface using a cotton swab. Microendoscopy images were taken from clinically normal cervix mucosa and from detected lesions. An image processing is performed to evaluate the cell nuclei morphology and the cytoplasm/nuclei ratio. The Pap smear results and the histology analyses are taken as gold standard for the diagnosis. Preliminary results in 5 patients demonstrated the potential use of our microscope at the clinical setting.

Surgical resection remains the primary curative intervention for cancer treatment. However, the occurrence of a residual tumor after resection is very common, leading to the recurrence of the disease and the need for re-resection. We develop a surgical Google Glass navigation system that combines near infrared fluorescent imaging and ultrasonography for intraoperative detection of sites of tumor and assessment of surgical resection boundaries, well as for guiding sentinel lymph node (SLN) mapping and biopsy. The system consists of a monochromatic CCD camera, a computer, a Google Glass wearable headset, an ultrasonic machine and an array of LED light sources. All the above components, except the Google Glass, are connected to a host computer by a USB or HDMI port. Wireless connection is established between the glass and the host computer for image acquisition and data transport tasks. A control program is written in C++ to call OpenCV functions for image calibration, processing and display. The technical feasibility of the system is tested in both tumor simulating phantoms and in a human subject. When the system is used for simulated phantom resection tasks, the tumor boundaries, invisible to the naked eye, can be clearly visualized with the surgical Google Glass navigation system. This system has also been used in an IRB approved protocol in a single patient during SLN mapping and biopsy in the First Affiliated Hospital of Anhui Medical University, demonstrating the ability to successfully localize and resect all apparent SLNs. In summary, our tumor simulating phantom and human subject studies have demonstrated the technical feasibility of successfully using the proposed goggle navigation system during cancer surgery.

A rise in the use of near-infrared (NIR) fluorescent dyes or intrinsic fluorescent markers for surgical guidance and tissue diagnosis has triggered the development of NIR fluorescence imaging systems. Because NIR wavelengths are invisible to the naked eye, instrumentation must allow surgeons to visualize areas of high fluorescence. Current NIR fluorescence imaging systems have limited ease-of-use because they display fluorescent information on remote display monitors that require surgeons to divert attention away from the patient to identify the location of tissue fluorescence. Furthermore, some systems lack simultaneous visible light imaging which provides valuable spatial context to fluorescence images. We have developed a novel, portable NIR fluorescence imaging approach for intraoperative surgical guidance that provides information for surgical navigation within the clinician’s line of sight. The system utilizes a NIR CMOS detector to collect excited NIR fluorescence from the surgical field. Tissues with NIR fluorescence are overlaid with visible light to provide information on tissue margins directly on the surgical field. In vitro studies have shown this versatile imaging system can be applied to applications with both extrinsic NIR contrast agents such as indocyanine green and weaker sources of biological fluorescence such as parathyroid gland tissue. This non-invasive, portable NIR fluorescence imaging system overlays an image directly on tissue, potentially allowing surgical decisions to be made quicker and with greater ease-of-use than current NIR fluorescence imaging systems.

Bacterial infection is one of the major factors contributing to the compromised healing in chronic wounds. Sometimes bacteria biofilms formed on the wound are more resistant than adherent bacteria. Cold atmosphere plasma (CAP) has already shown its potential in contact-free disinfection, blood coagulation, and wound healing. In this study, we integrated a multimodal imaging system with a portable CAP device for image-guided treatment of infected wound in vivo and evaluated the antimicrobial effect on Pseudomonas aeruginosa sample in vitro.15 ICR mice were divided into three groups for therapeutic experiments:(1) control group with no infection nor treatment (2) infection group without treatment (3) infection group with treatment. For each mouse, a three millimeters punch biopsy was created on the dorsal skin. Infection was induced by Staphylococcus aureus inoculation one day post-wounding. The treated group was subjected to CAP for 2 min daily till day 13. For each group, five fixed wounds’ oxygenation and blood perfusion were evaluated daily till day 13 by a multimodal imaging system that integrates a multispectral imaging module and a laser speckle imaging module. In the research of relationship between therapeutic depth and sterilization effect on P.aeruginosa in agarose, we found that the CAP-generated reactive species reached the depth of 26.7μm at 30s and 41.6μm at 60s for anti-bacterial effects. Image-guided CAP therapy can be potentially used to control infection and facilitate the healing process of infected wounds.

The significant increase of skin cancer occurring in the western world is attributed to longer sun expose during leisure time. For prevention, people should become aware of the risks of UV light exposure by showing skin damage and the protective effect of sunscreen with an UV camera. An UV awareness imaging system optimized for 365 nm (UV-A) was develop using consumer components being interactive, safe and mobile. A Sony NEX5t camera was adapted to full spectral range. In addition, UV transparent lenses and filters were selected based on spectral characteristics measured (Schott S8612 and Hoya U‐340 filters) to obtain the highest contrast for e.g. melanin spots and wrinkles on the skin. For uniform UV illumination, 2 facial tanner units were adapted with UV 365 nm black light fluorescent tubes. Safety of the UV illumination was determined relative to the sun and with absolute irradiance measurements at the working distance. A maximum exposure time over 15 minutes was calculate according the international safety standards. The UV camera was successfully demonstrated during the Dutch National Skin Cancer day and was well received by dermatologists and participating public. Especially, the 'black paint' effect putting sun screen on the face was dramatic and contributed to the awareness of regions on the face what are likely to be missed applying sunscreen. The UV imaging system shows to be promising for diagnostics and clinical studies in dermatology and potentially in other areas (dentistry and ophthalmology)

Polarized light has many applications in biomedical imaging. The interaction of a biological sample with polarized light reveals information about its composition, both structural and functional. For example, the polarimetry-derived metric of linear retardance (birefringence) is dependent on tissue structural organization (anisotropy) and can be used to diagnose myocardial infarct; circular birefringence (optical rotation) can measure glucose concentrations. The most comprehensive type of polarimetry analysis is to measure the Mueller matrix, a polarization transfer function that completely describes how a sample interacts with polarized light. To derive this 4x4 matrix it is necessary to observe how a tissue interacts with different polarizations. A well-suited approach for tissue polarimetry is to use photoelastic modulators (PEMs), which dynamically modulate the polarization of light. Previously, we have demonstrated a rapid time-gated Stokes imaging system that is capable of characterizing the state of polarized light (the Stokes vector) over a large field, after interacting with any turbid media. This was accomplished by synchronizing CCD camera acquisition times relative to two PEMs using a field-programmable gate array (FPGA). Here, we extend this technology to four PEMs, yielding a polarimetry system that is capable of rapidly measuring the complete sample Mueller matrix over a large field of view, with no moving parts and no beam steering. We describe the calibration procedure and evaluate the accuracy of the measurements. Results are shown for tissue-mimicking phantoms, as well as initial biological samples.

Pulmonary nodule could be identified by intraoperative fluorescence imaging system from systemic injection of indocyanine green (ICG) which achieves enhanced permeability and retention (EPR) effects. This study was performed to evaluate optimal injection time of ICG for detecting cancer during surgery in rabbit lung cancer model. VX2 carcinoma cell was injected in rabbit lung under fluoroscopic computed tomography-guidance. Solitary lung cancer was confirmed on positron emitting tomography with CT (PET/CT) 2 weeks after inoculation. ICG was administered intravenously and fluorescent intensity of lung tumor was measured using the custom-built intraoperative color and fluorescence merged imaging system (ICFIS) for 15 hours. Solitary lung cancer was resected through thoracoscopic version of ICFIS. ICG was observed in all animals. Because Lung has fast blood pulmonary circulation, Fluorescent signal showed maximum intensity earlier than previous studies in other organs. Fluorescent intensity showed maximum intensity within 6-9 hours in rabbit lung cancer. Overall, Fluorescent intensity decreased with increasing time, however, all tumors were detectable using fluorescent images until 12 hours. In conclusion, while there had been studies in other organs showed that optimal injection time was at least 24 hours before operation, this study showed shorter optimal injection time at lung cancer. Since fluorescent signal showed the maximum intensity within 6-9 hours, cancer resection could be performed during this time. This data informed us that optimal injection time of ICG should be evaluated in each different solid organ tumor for fluorescent image guided surgery.

There is no gold standard test for perfusion evaluation in surgery. Optical Imaging techniques are able to image tissue at high resolution and in real-time. Laser Speckle Contrast Imaging, Optical Coherence Tomography, Sidestream Darkfield and Incident Darkfield all use the interaction of light with tissue to create an image. To test their feasibility and explore validity in a controlled setting, we created a phantom with the optical properties of tissue and microvascular channels of 30-400 micrometer. With a Hamilton Syringe Pump we mimicked blood flow velocities of 0-20 mm/sec. Images of all different modalities at different blood flow velocities were compared in terms of imaging depth, resoluation and hemodynamic parameters.

Full-field optical coherence tomography (FFOCT) offers a non-invasive method of obtaining images of biological tissues at ultrahigh resolution (1µm in all 3 directions) approaching traditional histological sections. Previous clinical studies have shown the high efficiency of this imaging technique for the detection of cancer on various organs. This promises great potential of the technique for an ex-vivo quick analysis of surgical resections or biopsy specimens, in the aim to help the surgeon/radiologist decide on the course of action. Here we will present some of the latest technical developments on a FFOCT system which can produce 1cm2 images with 1 µm resolution in 1 minute. Larger samples, up to 50mm diameter, can also be imaged. Details on the large sample handling, high-speed image acquisition, optimized scanning, and accelerated GPU tiles stitching will be given. Results on the clinical applications for breast, urology, and digestive tissues will also be given. They highlight the relevance of the system characteristics for the detection of cancer on ex-vivo specimens. FFOCT now appears clearly as a very fast and non-destructive imaging technique that provides a quick assessment of the tissue morphology. With the benefit of both new technical developments and clinical validation, it turned into a mature technique to be implemented in the clinical environment. In particular, the technique holds potential for the fast ex-vivo analysis of excision margins or biopsies in the operating room.

Despite the improvements in early cancer diagnosis, adequate diagnostic tools for early staging of bladder cancer tumors are lacking [1]. MEMS-probes based on optical coherence tomography (OCT) provide cross-sectional imaging with a high-spatial resolution at a high-imaging speed, improving visualization of cancerous tissue [2-3]. Additionally, studies show that the measurement of localized attenuation coefficient allows discrimination between healthy and cancerous tissue [4]. We have designed a new miniaturized MEMS-probe based on OCT that will optimize early diagnosis by improving functional visualization of suspicious lesions in bladder. During the optical design phase of the probe, we have studied the effect of the numerical aperture (NA) on the OCT signal attenuation. For this study, we have employed an InnerVision Santec OCT system with several numerical apertures (25mm, 40mm, 60mm, 100mm, 150mm and 200mm using achromatic lenses). The change in attenuation coefficient was studied using 15 dilutions of intralipid ranging between 6*10-5 volume% and 20 volume%. We obtained the attenuation coefficient from the OCT images at several fixed positions of the focuses using established OCT models (e.g. single scattering with known confocal point spread function (PSF) [5] and multiple scattering using the Extended Huygens Fresnel model [6]). As a result, a non-linear increase of the scattering coefficient as a function of intralipid concentration (due to dependent scattering) was obtained for all numerical apertures. For all intralipid samples, the measured attenuation coefficient decreased with a decrease in NA. Our results suggest a non-negligible influence of the NA on the measured attenuation coefficient. [1] Khochikar MV. Rationale for an early detection program for bladder cancer. Indian J Urol 2011 Apr-Jun; 27(2): 218–225. [2] Sun J and Xie H. Review Article MEMS-Based Endoscopic Optical Coherence Tomography. IJO 2011, Article ID 825629, 12 pages. doi:10.1155/2011/825629. [3] Jung W and Boppart S. Optical coherence tomography for rapid tissue screening and directed histological sectioning. Anal Cell Pathol (Amst). 2012; 35(3): 129–143. [4] R. Wessels et al. Optical coherence tomography in vulvar intraepithelial neoplasia. J Biomed Opt 2012 Nov; 17(11): 116022. [5] Faber D, van der Meer F, Aalders M, van Leeuwen T. Quantitative measurement of attenuation coefficients of weakly scattering media using optical coherence tomography. OPT EXPRESS 2004; 12 (19): 4353-43. [6] Thrane L, Yura HT, and Andersen PE. Analysis of optical coherence tomography systems based on the extended Huygens–Fresnel principle. JOSA 2000; 17(3): 484-490.

Optical Coherence Tomography (OCT) can discriminate morphological tissue features important for oral cancer detection such as the presence or absence of basement membrane and epithelial thickness. We previously reported an OCT system employing a rotary-pullback catheter capable of in vivo, rapid, wide-field (up to 90 x 2.5mm2) imaging in the oral cavity. Due to the size and complexity of these OCT data sets, rapid automated image processing software that immediately displays important tissue features is required to facilitate prompt bed-side clinical decisions. We present an automated segmentation algorithm capable of detecting the epithelial surface and basement membrane in 3D OCT images of the oral cavity. The algorithm was trained using volumetric OCT data acquired in vivo from a variety of tissue types and histology-confirmed pathologies spanning normal through cancer (8 sites, 21 patients). The algorithm was validated using a second dataset of similar size and tissue diversity. We demonstrate application of the algorithm to an entire OCT volume to map epithelial thickness, and detection of the basement membrane, over the tissue surface. These maps may be clinically useful for delineating pre-surgical tumor margins, or for biopsy site guidance.

Brain needle biopsy (BNB) is performed to collect tissue when precise neuropathological diagnosis is required to provide information about tumor type, grade, and growth patterns. The principal risks associated with this procedure are intracranial hemorrhage (due to clipping blood vessels during tissue extraction), incorrect tumor typing/grading due to non-representative or non-diagnostic samples (e.g. necrotic tissue), and missing the lesion. We present an innovative device using sub-diffuse optical tomography to detect blood vessels and Raman spectroscopy to detect molecular differences between tissue types, in order to reduce the risks of misdiagnosis, incorrect tumour grading, and non-diagnostic samples. The needle probe integrates optical fibers directly onto the external cannula of a commercial BNB needle, and can perform measurements for both optical techniques through the same fibers. This integrated optical spectroscopy system uses diffuse reflectance signals to perform a 360-degree reconstruction of the tissue adjacent to the biopsy needle, based on the optical contrast associated with hemoglobin light absorption, thereby localizing blood vessels. Raman spectra measurements are also performed interstitially for tissue characterization. A detailed sensitivity of the system is presented to demonstrate that it can detect absorbers with diameters <300 µm located up to ∼2 mm from the biopsy needle core, for bulk optical properties consistent with brain tissue. Results from animal experiments are presented to validate blood vessel detection and Raman spectrum measurement without disruption of the surgical workflow. We also present phantom measurements of Raman spectra with the needle probe and a comparison with a clinically validated Raman spectroscopy probe.

While there are a plethora of in-vivo spectroscopic techniques that have demonstrated the ability to detect a number of diseases in research trials, very few techniques have successfully become a fully realized clinical technology. This is primarily due to the stringent demands on a clinical device for widespread implementation. Some of these demands include: simple operation requiring minimal or no training, safe for in-vivo patient use, no disruption to normal clinic workflow, tracking of system performance, warning for measurement abnormality, and meeting all FDA guidelines for medical use. Previously, our group developed a fiber optic probe-based optical sensing technique known as low-coherence enhanced backscattering spectroscopy (LEBS) to quantify tissue ultrastructure in-vivo. Now we have developed this technique for the application of prescreening patients for colonoscopy in a primary care (PC) clinical setting. To meet the stringent requirements for a viable medical device used in a PC clinical setting, we developed several novel components including an automated calibration tool, optical contact sensor for signal acquisition, and a contamination sensor to identify measurements which have been affected by debris. The end result is a state-of-the-art medical device that can be realistically used by a PC physician to assess a person’s risk for harboring colorectal precancerous lesions. The pilot study of this system shows great promise with excellent stability and accuracy in identifying high-risk patients. While this system has been designed and optimized for our specific application, the system and design concepts are universal to most in-vivo fiber optic based spectroscopic techniques.

The mechanism of most medical endoscopes is based on the interaction between light and biological tissue, inclusive of absorption, elastic scattering and fluorescence. In essence, the metrics of those interactions are obtained from the fundamental properties of light as an electro-magnetic waves, namely, the radiation intensity and wavelength. As another fundamental property of light, polarisation can not only reveal tissue scattering and absorption information from a different perspective, but is also able to provide a fresh insight into directional tissue birefringence properties induced by birefringent compositions and anisotropic fibrous structures, such as collagen, elastin, muscle fibre, etc at the same time. Here we demonstrate a low cost high definition Muller polarimetric endoscope with minimal alteration of a rigid endoscope. By imaging birefringent tissue mimicking phantoms and a porcine bladder, we show that this novel endoscopic imaging modality is able to provide different information of interest from unpolarised endoscopic imaging, including linear depolarization, circular depolarization, birefringence, optic axis orientation and dichroism. This endoscope can potentially be employed for better tissue visualisation and benefit endoscopic investigations and intra-operative guidance.

Mueller matrix imaging along with polar decomposition method was employed for the colonic adenocarcinoma detection by polarized light in the near-infrared spectral range (700–1100 nm). A high-speed (<5s) Muller matrix imaging system with dual-rotating waveplates was developed. 16 (4 by 4) full Mueller matrices of the colonic tissues (i.e., normal and caner) were acquired. Polar decomposition was further implemented on the 16 images to derive the diattentuation, depolarization, and the retardance images. The decomposed images showed clear margin between the normal and adenocarcinomaous colon tissue samples. The work shows the potential of near-infrared Mueller matrix imaging for the early diagnosis and detection of malignant lesions in the colon.

Near-infrared Spectroscopy (NIRS) is a noninvasive technique to monitor tissue oxygenation dynamics and microcirculation. Because of the advantage, sepsis has been investigated with NIRS for many years including severe sepsis and sepsis shock. However, it is necessary to involve a physical method, which is often venous occlusion to change hemodynamics. There are some drawbacks of vessel-occlusion method that include skin contact, uncomfortable and microcirculation block of patients. In order to improve the user experience from venous occlusion, we developed a new method which is far-infrared illumination. In this study, we investigated sepsis patient with far-infrared illumination and NIRS. The result release that this method could recognize severe sepsis and sepsis shock with good user experience. We hope this method can be used for clinical diagnosis of other peripheral arterial disease.

Skin blood microcirculation and the metabolism activity of tissue were examined on the patients with type 2 diabetes. Laser Doppler flowmetry (LDF) with 1064 nm laser light source and fluorescence spectroscopy (FS) with excitation light of 365 nm and 450 nm have been used to monitor the blood perfusion and the content of coenzymes NADH and FAD. Concluding, the proposed combined LDF and tissue FS approach allows to identify the significant violations in the blood microcirculation and metabolic activity for type 2 diabetes patients.

Optical coherence tomography (OCT) is a noninvasive imaging tool for visualizing cross-sectional images of biological tissues on a microscale. Various microelectromechanical system (MEMS) techniques have been applied to OCT for endoscopic catheters and handheld probes. Despite having several advantages such as compact sizes and high speeds for real-time imaging, the complexities of the fabrication processes and relatively high costs were bottlenecks for fast clinical translation and commercialization of the earlier MEMS scanners. To overcome these issues, we developed a 2-axis polydimethylsiloxane (PDMS)-based electromagnetic MEMS scanning mirror based on flexible, cost-effective, and handleable PDMS. The size of this MEMS scanner was 15 × 15 × 15 mm³. To realize the characteristics of the scanner, we obtained the DC/AC responses and scanning patterns. The measured maximum scanning angles were 16.6° and 11.6° along the X and Y axes, respectively. The resonance frequencies were 82 and 57 Hz along the X and Y axes, respectively. The scanning patterns (raster and Lissajous scan patterns) are also demonstrated by controlling the frequency and amplitude. Finally, we showed the in vivo 2D-OCT images of human fingers by using a spectral domain OCT system with a PDMSbased MEMS scanning mirror. We then reconstructed the 3D images of human fingers. The obtained field of view was 8 × 8 mm². The PDMS-based MEMS scanning mirror has the potential to combine other optical modalities and be widely used in preclinical and clinical translation research.

Diffuse correlation spectroscopy (DCS) is a technique which enables powerful and robust non-invasive optical studies of tissue micro-circulation and vascular blood flow. The technique amounts to autocorrelation analysis of coherent photons after their migration through moving scatterers and subsequent collection by single-mode optical fibers. A primary cost driver of DCS instruments are the commercial hardware-based correlators, limiting the proliferation of multi-channel instruments for validation of perfusion analysis as a clinical diagnostic metric. We present the development of a low-cost scalable correlator enabled by microchip-based time-tagging, and a software-based multi-tau data analysis method. We will discuss the capabilities of the instrument as well as the implementation and validation of 2- and 8-channel systems built for live animal and pre-clinical settings.

The schwannomas is a tumour of the tissue that covers nerves, called the nerve sheath. Schwannomas are often benign tumors of the Schwan cells, which are the principal glia of the peripheral nervous system (PNS). Preoperative diagnosis of this lesion usually is difficult, therefore, new techniques are being studied as pre surgical evaluation. Among these, Raman spectroscopy, that enables the biochemical identification of the tissue analyzed by their optical properties, may be used as a tool for schwannomas diagnosis. The aim of this study was to discriminate between normal nervous tissue and schwannoma through the confocal Raman spectroscopy and Raman optical fiber-based techniques combined with immunohistochemical analysis. Twenty spectra were analyzed from a normal nerve tissue sample (10) and schwannoma (10) by Holospec f / 1.8 (Kayser Optical Systems) coupled to an optical fiber with a 785nm laser line source. The data were pre-processed and vector normalized. The average analysis and standard deviation was performed associated with cluster analysis. AML, 1A4, CD34, Desmin and S-100 protein markers were used for immunohistochemical analysis. Immunohistochemical analysis was positive only for protein S-100 marker which confirmed the neural schwanomma originality. The immunohistochemistry analysis were important to determine the source of the injury, whereas Raman spectroscopy were able to differentiated tissues types indicating important biochemical changes between normal and benign neoplasia.

Functional gastrointestinal disorders (FGID) are the most common gastrointestinal disorders. The term ”functional” is generally applied to disorders where there are no structural abnormalities. Gastrointestinal dysmotility is one of the several mechanisms that have been proposed for the pathogenesis of FGID and is usually examined by manometry, a pressure test. There have been no attempts to examine the gastrointestinal dysmotility by endoscopy. We have proposed an imaging system for the assessment of gastric motility using a three-dimensional endoscope. After we newly developed a threedimensional endoscope and constructed a wave simulated model, we established a method of extracting three-dimensional contraction waves derived from a three-dimensional profile of the wave simulated model obtained with the endoscope. In the study, the endoscope and the wave simulated model were fixed to the ground. However, in a clinical setting, it is hard for endoscopists to keep the endoscope still. Moreover, stomach moves under the influence of breathing. Thus, three-dimensional registration of the position between the endoscope and the gastric wall is necessary for the accurate assessment of gastrointestinal motility. In this paper, we propose a motion compensation method using three-dimensional scene flow. The scene flow of the feature point calculated by obtained images in a time series enables the three-dimensional registration of the position between the endoscope and the gastric wall. We confirmed the validity of a proposed method first by a known-movement object and then by a wave simulated model.

Herein, a finger-free wrist-worn pulse oximeter is presented. This device allows patients to measure blood oxygen level and pulse rate without hindering their normal finger movement. This wrist-worn pulse oximeter is built with a reflectance oximetry sensor, which consists of light emitting diodes and photodiode light detectors located side by side. This reflectance oximetry sensor is covered with an optical element with micro structured surface. This micro structured optical element is designed to modulate photon propagation beneath the skin tissue so that the photoplethysmogram signals of reflected lights or backscattered lights detected by the photodetector are therefore enhanced.

Stereoscopic retinal image can effectively help doctors. Most of stereo imaging surgical microscopes are based on dual optical channels and benefit from dual cameras in which left and right cameras capture corresponding left and right eye views. This study developed a single-channel stereoscopic retinal imaging modality based on a transparent rotating deflector (TRD). Two different viewing angles are generated by imaging through the TRD which is mounted on a motor synchronized with a camera and is placed in single optical channel. Because of the function of objective lens in the imaging modality which generate stereo-image from an object at its focal point, and according to eye structure, the optical set up of the imaging modality can compatible for retinal imaging when the cornea and eye lens are engaged in objective lens.

Clinical shock-monitoring mainly depends on measuring oxygen saturations from SVC blood samples invasively. The golden standard indicator is the central internal jugular vein oxygenation (SjvO₂). Using near-infrared spectroscopy (NIRS) also can monitor shock in some papers published, but there is no discussion about which oxygen saturation (cerebral venous oxygen saturation, ScvO₂; tissue oxygen saturation of internal jugular area; tissue oxygen saturation of extremities areas) can monitor shock patient more sensitively and accurately. The purpose of this paper is to examine which one is most effective. In order to discuss the problem, we continuously detected 56 critical patients who may be into shock state using NIRS oximeter at prefrontal, internal jugular vein area and forearm, and chose 24 patients who were into shock and then out of shock from the 56 critical patients. Combined with the patients’ condition, the pulse oxygen saturation is most sensitively to monitoring shock than the others, and the internal jugular vein area oxygen saturation is most effective.

This article introduces a novel method to estimate oxygen saturation of the internal jugular vein blood (SjvO2) by using Near Infrared spectroscopy (NIRS). The different positions of patients can affect the cross-sectional area (CSA) of the internal jugular vein (IJV), in other words, it causes the sectional change of the IJV blood volume. When lying position of patients, the CSA is larger than that keeping upper body 80 degree, and the CSA can compute quantitatively by the use of ultrasound and digital image processing methods. The entire method consist of constructing different position of patient (upper body rotation 0 and 80 degree), comparing the light absorption changes. SjvO₂ has been determined from light absorption measurements in two wavelength, before and after the position changes. The method has been applied to the vertical area over the IJV of 11 patients who were placed a central venous catheter into a large vein in the neck for medical uses, using wavelength of 735 and 850 nm. At last, comparing the SjvO_2NIRS which measured by NIRS noninvasively with SjvO_2IJVBG which was quantified using a whole blood gas analyzer, we found there were some certain relativity. The results were influenced by vascular depth greatly.