This PDF file contains the front matter associated with SPIE Proceedings Volume 10411 including the Title Page, Copyright information, Table of Contents, Introduction, and Conference Committee listing.

Fluorescence lifetime imaging microscopy (FLIM) is a promising non-invasive highly sensitive technique for probing multiple physiological and physicochemical parameters in living cells and tissues. The present study is focused on the investigation of bioenergetics and microscopic viscosity of cultured cancer cells and animal tumors using FLIM during natural growth and chemotherapy. Fluorescence lifetime measurements of the metabolic cofactor NAD(P)H revealed a decrease of the relative amplitude of free NAD(P)H after cisplatin treatment, indicating a change towards a more oxidative metabolic state. Microviscosity mapping performed with the use of fluorescent molecular rotor BODIPY-2 showed a pronounced increase in the plasma membrane viscosity in cancer cells exposed to cisplatin. Although biochemical mechanisms underlying the metabolic and viscosity alterations during chemotherapy have yet to be clarified, our data suggest that the cisplatin-induced changes in cellular metabolism and membrane viscosity play a role in the cytotoxicity of the drug. The results of the study contribute to an understanding of mechanisms of cisplatin action and will be useful for development new approach for assessing response to a therapy.

Raman microspectroscopy is well suited for readily revealing information about bio-samples. As such, this technique has been applied to a wide range of areas, especially in bio-medical diagnosis. However, bio-samples typically suffer from low Raman signal level due to the nature of inelastic scattering of photons, To achieve a decent signal level, usually a high numerical aperture is employed. One drawback with these objectives is that their working distance is very short. In many cases of clinic diagnosis, a long working distance is always desired which limits the usage of these objectives. We propose a practical solution to this problem by enhancing the Raman/fluorescence signal by a parabolic reflector. On one hand, the high signal level is achieved by the large solid angle of collection of the parabolic reflector. On the other hand, the long working distance is guaranteed by the novel design of our microscope. The enhancement-capability is demonstrated through five types of samples among which we found the method is most applicable for turbid samples.

Linear polarization-in, polarization-out second-harmonic generation microscopy was used to study the effect of Photodynamic therapy treatment on enhancing the healing of femur fracture by investigating the ultrastructure of collagen as a major component of bone matrix.

Velocity signals acquired with a Laser Doppler Vibrometer on the thorax (Optical Vibrocardiography) contain important information, which have a relation to cardiovascular parameters and cardiovascular diseases. The acquired signal results in a superimposition of vibrations originated from different sources of the human body. Since we study the vibration generated by the heart to reliably detect a characteristic time interval corresponding to the PR interval in the ECG, these disturbance have to be removed by filtering. Moreover, the Laser Doppler Vibrometer measures only in the direction of the laser beam and, thus, the velocity signal is only a projection of the tridimensional movement of the thorax. This work presents an analysis of the influences of the filters and of the measurement direction on the characteristic time interval in Vibrocardiographic signals. Our analysis results in recommended settings for filters and we demonstrate that reliable detection of vibrocardiographic parameters is possible within an angle deviation of 30° in respect to the perpendicular irradiation on the front side of the subject.

Despite the wide clinical uses of pulse-oximetry, the precise nature of the light-tissue interaction underneath the technique is not clearly understood. A heterogeneous opto-anatomical model is presented to describe the optical path in pulse oximetry.

We present the first broadband (600-1100 nm) diffuse optical characterization of thyroglobulin and tyrosine, which are thyroid-specific tissue constituents. In-vivo measurements at the thyroid region enabled their quantification for functional and diagnostic applications.

In this study was evaluated the photoaging of patients' skins by the processing of RGB images acquired with an optical system based on a smartphone. Two groups were approached: a younger and an older.

Spectra computed from multispectral images of murine models of Rheumatoid Arthritis show a characteristic decrease in reflectance within the 600-800nm region which is indicative of the reduction in blood oxygenation and is consistent with hypoxia.

A multispectral endoscope capable of simultaneous reflectance and fluorescence imaging was developed based on spectrally resolved detector arrays. The endoscope can simultaneous image and unmix oxy/deoxygenated blood and two fluorescent dyes.

In this paper, the multi-spectral endogenous fluorescence imaging was implemented for bacterial differentiation. The fluorescence imaging was performed using a digital camera equipped with a set of visual bandpass filters. Narrowband 365 nm ultraviolet radiation passed through a beam homogenizer was used to excite the sample fluorescence. In order to increase a signal-to-noise ratio and suppress a non-fluorescence background in images, the intensity of the UV excitation was modulated using a mechanical chopper. The principal components were introduced for differentiating the samples of bacteria based on the multi-spectral endogenous fluorescence images.

We show the feasibility of using an intraoperative spectroscopic device to identify tumors margins during glioma resection. The collected fluorescence spectra is fitted with two reference spectra of PpIX and the contribution of each spectrum enables to overcome the sensitivity of current techniques by seeing tumor margins and low grade gliomas.

Second harmonic generation (SHG) is a microscopic technique applicable to a broad spectrum of biological and medical imaging due to its excellent photostability, high signal-to-noise ratio (SNR) and narrow emission profile. Current SHG microscopy techniques rely on two main contrast modalities. These are endogenous SHG generated by tissue structures, which is clinically relevant but cannot be targeted to another location, or SHG nanoprobes, inorganic nanocrystals that can be directed to proteins and cells of interest, but cannot be applied for clinical imaging due to their chemical composition. Here we analyzed SHG signal generated by large-scale peptide assemblies. Our results show the sequence of peptides play an important role on both the morphology and SHG signal of the peptide assemblies. Changing peptide sequence allows confinement of large number of peptides to smaller voxels, generating intense SHG signal. With miniaturization of these peptides and their proper functionalization strategies, such bioinspired nanoparticles would emerge as valuable tools for clinical imaging.

An optical fiber probe for multimodal spectroscopy was designed, developed and used for tissue diagnostics. The probe, based on a fiber bundle with optical fibers of various size and properties, allows performing spectroscopic measurements with different techniques, including fluorescence, Raman, and diffuse reflectance, using the same probe. Two visible laser diodes were used for fluorescence spectroscopy, a laser diode emitting in the NIR was used for Raman spectroscopy, and a fiber-coupled halogen lamp for diffuse reflectance. The developed probe was successfully employed for diagnostic purposes on various tissues, including brain and bladder. In particular, the device allowed discriminating healthy tissue from both tumor and dysplastic tissue as well as to perform tumor grading. The diagnostic capabilities of the method, determined using a cross-validation method with a leave-one-out approach, demonstrated high sensitivity and specificity for all the examined samples, as well as a good agreement with histopathological examination performed on the same samples. The obtained results demonstrated that the multimodal approach is crucial for improving diagnostic capabilities with respect to what can be obtained from individual techniques. The experimental setup presented here can improve diagnostic capabilities on a broad range of tissues and has the potential of being used clinically for guiding surgical resection in the near future.

In-vivo fluorescently labelled drug (bevacizumab) breast cancer specimen where obtained from patients. We propose a new structured method to determine the optimal classification threshold in targeted fluorescence intra-operative imaging.

We present our recent work using optical imaging and spectroscopy to investigate changes associated with oxygenation and metabolism in radiation-resistant and sensitive tumors.

We present a computer model of intravital excitation and external fluorescence detection in the murine lungs validated with a three-dimensional lung tissue phantom. The model is applied to optical detection of pulmonary tuberculosis infection.

Blood flow monitoring during rhythm exercising is very important for sports medicine and muscle dieases. Diffuse correlation spectroscopy(DCS) is a relative new invasive way to monitor blood flow but suffering from muscle fiber motion. In this study we focus on how to remove exercise driven artifacts and obtain accurate estimates of the increase in blood flow from exercise. Using a novel fast software correlator, we measured blood flow in forearm flexor muscles of N=2 healthy adults during handgrip exercise, at a sampling rate of 20 Hz. Combining the blood flow and acceleration data, we resolved the motion artifact in the DCS signal induced by muscle fiber motion, and isolated the blood flow component of the signal from the motion artifact. The results show that muscle fiber motion strongly affects the DCS signal, and if not accounted for, will result in an overestimate of blood flow more than ~1000%. Our measurements indicate rapid dilation of arterioles following exercise onset, which enabled blood flow to increase to a plateau of ~200% in ~10s. The blood flow also rapidly recovered to baseline following exercise in ~10s. Finally, preliminary results on the dependence of blood flow from exercise intensity changes will be discussed.

Despite recent advances in fluorescence imaging, standardization of systems remains an unmet need. We developed a new comprehensive phantom that resolves multiple system parameters simultaneously and could be used for system performance comparison.

There is a pressing clinical need to provide image guidance during surgery. Currently, assessment of tissue that needs to be resected or avoided is performed subjectively leading to a large number of failures, patient morbidity and increased healthcare cost. Because near-infrared (NIR) optical imaging is safe, does not require contact, and can provide relatively deep information (several mm), it offers unparalleled capabilities for providing image guidance during surgery. In this work, we introduce a novel concept that enables the quantitative imaging of endogenous molecular information over large fields-of-view. Because this concept can be implemented in real-time, it is amenable to provide video-rate endogenous information during surgery.

The relative occurrence of volatile organic compounds in the human respiratory gas is disease-specific (ppb range). A prototype of a gas analysing device using two tuneable laser systems, an OPO-laser (2.5 to 10 μm) and a CO₂-laser (9 to 11 μm), and an optoacoustic measurement cell was developed to detect concentrations in the ppb range. The sensitivity and resolution of the system was determined by test gas measurements, measuring ethylene and sulfur hexafluoride with the CO₂-laser and butane with the OPO-laser. System sensitivity found to be 13 ppb for sulfur hexafluoride, 17 ppb for ethylene and <10 ppb for butane, with a resolution of 50 ppb at minimum for sulfur hexafluoride. Respiratory gas samples of 8 healthy volunteers were investigated by irradiation with 17 laser lines of the CO₂-laser. Several of those lines overlap with strong absorption bands of ammonia. As it is known that ammonia concentration increases by age a separation of people <35 und >35 was striven for. To evaluate the data the first seven gas samples were used to train a discriminant analysis algorithm. The eighth subject was then assigned correctly to the group >35 years with the age of 49 years.

Urinary tract infection (UTI) is one of the major clinical problems known to mankind, especially among adult women. Conventional methods for identification of UTI causing bacteria are time consuming and expensive. Therefore, a rapid and cost-effective method is desired. In the present study, five bacteria (one Gram-positive and four Gram-negative), most commonly known to cause UTI, have been identified and classified using micro-Raman spectroscopy combined with principal component analysis (PCA).

Significant contrast in visible wavelength Mueller matrix images for healthy and pre-cancerous regions of excised cervical tissue is shown. A novel classification algorithm is used to compute a test statistic from a small patient population.

This contribution presents a fast global adjustment scheme exploiting SURF descriptor locations for constructing large skin mosaics. Precision in pairwise image registration is well-preserved while significantly reducing the global mosaicing error.

Skin lesion segmentation is a complex step for dermoscopy pathological diagnosis. Kernel density estimation is proposed as a segmentation technique based on the statistic distribution of color intensities in the lesion and non-lesion regions.

In this paper, a handheld contact probe based on sapphire shaped crystal was developed for the intraoperative optical diagnosis and aspiration of malignant brain tissue combined with the laser hemostasis. Such a favorable combination of several functions in a single instrument significantly increases its clinical relevance. It makes possible highly-accurate real-time detection and removal of either large-scale malignancies or even separate invasive cancer cells. The proposed neuroprobe was integrated into the clinical neurosurgical workflow for the intraoperative fluorescence identification and removal of malignant tissues of the brain.

Using multidetector computed tomography, thicknesses of bone squame and soft tissues of human head were assessed. MC simulation revealed impropriety of source-detector separation distances for 3 oximeters, which can cause extracerebral contamination.

We have developed the method for imaging blood flow and blood concentration change by using laser speckle in fiber illumination. We experimentally discuss the relationship of blood occlusion condition and individual response of blood concentration change measured by the method.

In present study, remote photoplethysmography signals acquired by ultra-low noise camera and conventional camera were compared during different skin microcirculation provocation tests. The aim of work was to reveal how much of camera dynamic range and noise contribute to blood perfusion signal quality. Results demonstrate comparable capabilities of both cameras for skin perfusion monitoring.

Multimodal OCT is a promising tool for monitoring of individual tumor response to antitumor therapies. The changes of tumor cells, connective tissue, microcirculation and stiffness can be estimated simultaneously in real time with high resolution.

Autofluorescence spectroscopy offer noninvasive and promising tools for the detection of alternations biochemical compositions of tissues and cells, in presence of disease. They have the added advantage of being highly objective due to the fact that diagnostic evaluation is by statistical methods, eliminating errors from lack of experience, fatigue factor, and subjectivity of visual perceptions. The present research work involves in designing and assembling of a low cost, miniature oral cancer screening device with for routine clinical applications. A miniature system was designed and assembled with much smaller and cost-effective components like compact light source and miniature spectrometer, in a hand-held unit configuration. The performance of the system was evaluated using animal -mouse- SCC model. The current system can be used in handheld operation, which makes it very useful for many applications like, screening of squamous cell carcinoma susceptible population.

Surgical repair of the mitral valve complex presents high mortality rates, strongly dependent on the surgical procedure. Intensity and polarization sensitive OCT are explored as a feasible degradation inspection method for rheumatic and degenerative chords.

Using diffuse optical spectroscopy the level of oxygenation and hemoglobin concentration in experimental tumor in comparison with normal muscle tissue of mice have been studied. Subcutaneously growing SKBR-3 was used as a tumor model. Continuous wave fiber probe diffuse optical spectroscopy system was employed. Optical properties extraction approach was based on diffusion approximation. Decreased blood oxygen saturation level and increased total hemoglobin content were demonstrated in the neoplasm. The main reason of such differences between tumor and norm was significant elevation of deoxyhemoglobin concentration in SKBR-3. The method can be useful for diagnosis of tumors as well as for study of blood flow parameters of tumor models with different angiogenic properties.

We combined Raman micro-spectroscopy and machine learning techniques to develop a classification model based on a well-established non-alcoholic steatohepatitis (NASH) mouse model, using spectrum pre-processing, biochemical component analysis (BCA) and logistic regression.