The concept of using diffuse reflectance spectroscopy to distinguish intraoperatively between pediatric brain tumors and normal brain parenchyma at the edge of resection cavities is evaluated using an in vivo human study. Diffuse reflectance spectra are acquired from normal and tumorous brain areas of 12 pediatric patients during their tumor resection procedures, using a spectroscopic system with a handheld optical probe. A total of 400 spectra are acquired at the rate of 33 Hz from a single investigated site, from which the mean spectrum and the standard deviation are calculated. The mean diffuse reflectance spectra collected are divided into the normal and the tumorous categories in accordance with their corresponding results of histological analysis. Statistical methods are used to identify those spectral features that effectively separated the two tissue categories, and to quantify the spectral variations induced by the motion of the handheld probe during a single spectral acquisition procedure. The results show that diffuse reflectance spectral intensities between 600 and 800 nm are effective in terms of differentiating normal cortex from brain tumors. Furthermore, probe movements induce large variations in spectral intensities (i.e., larger standard deviation) between 400 and 600 nm.
KEYWORDS: Brain, Tissue optics, Diffuse reflectance spectroscopy, Epilepsy, Data analysis, Tissues, Tumors, Control systems, Statistical analysis, In vivo imaging
This research investigated the feasibility of using time-dependent diffuse reflectance spectroscopy to differentiate
pediatric epileptic brain tissue from normal brain tissue. The optical spectroscopic technique monitored the dynamic
optical properties of the cerebral cortex that are associated with its physiological, morphological, and compositional
characteristics. Due to the transient irregular epileptic discharge activity within the epileptic brain tissue it was
hypothesized that the lesion would express abnormal dynamic optical behavior that would alter normal dynamic
behavior. Thirteen pediatric epilepsy patients and seven pediatric brain tumor patients (normal controls) were recruited
for this clinical study. Dynamic optical properties were obtained from the cortical surface intraoperatively using a timedependent
diffuse reflectance spectroscopy system. This system consisted of a fiber-optic probe, a tungsten-halogen light
source, and a spectrophotometer. It acquired diffuse reflectance spectra with a spectral range of 204 nm to 932 nm at a
rate of 33 spectra per second for approximately 12 seconds. Biopsy samples were taken from electrophysiologically
abnormal cortex and evaluated by a neuropathologist, which served as a gold standard for lesion classification. For data
analysis, spectral intensity changes of diffuse reflectance in the time domain at two different wavelengths from each
investigated site were compared. Negative correlation segment, defined by the periods where the intensity changes at the
two wavelengths were opposite in their slope polarity, were extracted. The total duration of negative correlation, referred
to as the "negative correlation time index", was calculated by integrating the negative correlation segments. The negative
correlation time indices from all investigated sites were sub-grouped according to the corresponding histological
classifications. The difference between the mean indices of two subgroups was evaluated by standard t-test. These
comparison and calculation procedures were carried out for all possible wavelength combinations between 400 nm and
800 nm with 2 nm increments. The positive group consisted of seven pathologically abnormal test sites, and the negative
group consisted of 13 normal test sites from non-epileptic tumor patients. A standard t-test showed significant difference
between negative correlation time indices from the two groups at the wavelength combinations of 700-760 nm versus
550-580 nm. An empirical discrimination algorithm based on the negative correlation time indices in this range produced
100% sensitivity and 85% specificity. Based on these results time-dependent diffuse reflectance spectroscopy with
optimized data analysis methods differentiates epileptic brain tissue from normal brain tissue adequately, therefore can
be utilized for surgical guidance, and may enhance the surgical outcome of pediatric epilepsy surgery.
Optical spectroscopy for in vivo tissue diagnosis is performed traditionally in a static manner; a snap shot of the tissue
biochemical and morphological characteristics is captured through the interaction between light and the tissue. This
approach does not capture the dynamic nature of a living organ, which is critical to the studies of brain disorders such as
epilepsy. Therefore, a time-dependent diffuse reflectance spectroscopy system with a fiber-optic probe was designed
and developed. The system was designed to acquire broadband diffuse reflectance spectra (240 ~ 932 nm) at an
acquisition rate of 33 Hz. The broadband spectral acquisition feature allows simultaneous monitoring of various
physiological characteristics of tissues. The utility of such a system in guiding pediatric epilepsy surgery was tested in a
pilot clinical study including 13 epilepsy patients and seven brain tumor patients. The control patients were children
undergoing suregery for brain tumors in which measurements were taken from normal brain exposed during the surgery.
Diffuse reflectance spectra were acquired for 12 seconds from various parts of the brain of the patients during surgery.
Recorded spectra were processed and analyzed in both spectral and time domains to gain insights into the dynamic
changes in, for example, hemodynamics of the investigated brain tissue. One finding from this pilot study is that
unsynchronized alterations in local blood oxygenation and local blood volume were observed in epileptogenic cortex.
These study results suggest the advantage of using a time-dependent diffuse reflectance spectroscopy system to study
epileptogenic brain in vivo.
We report results of a study demonstrating that swept-source spectral interferometry and spectral phase analysis may be
applied to determine quantitatively the concentration of glucose in aqueous solutions. The optical system incorporates a
frequency swept laser and fiber-based common-path interferometer and provides a compact, stable experimental system.
Spectral phase analysis includes a fast Lomb-Scargle algorithm and a multitaper spectral estimation method that allows
for time-frequency analysis. The results show highly sensitive and accurate determination of glucose concentration in
solutions with 0.54 mM of resolution and 0.999 correlation coefficient. Swept-source spectral interferometry with
spectral phase analysis is a promising methodology for further development of a convenient glucose-sensing method
with superior sensitive and accuracy.
Speckle in optical coherence tomography (OCT) images originates in the high spatial coherence of incident light that enables interference of light backscattered from spatially heterogenous tissue specimens. We report results of a numerical simulation and an experiment to test speckle reduction using a partially spatially coherent source. A Gaussian-Schell model for a partially spatially coherent source is used in the OCT simulation. For the experiment, such a source was generated by a spatially coherent boardband light source and a multimode fiber. The advantage of using a multimode fiber in combination with a broadband source is the large number of photons per coherence volume. To illustrate speckle reduction with a partially spatially coherent source, we record low-coherence interferograms of a scattering surface using single-mode and multimode source fibers. Interferograms recorded using a single-mode source fiber are indicative of those observed using conventional OCT. Speckle in OCT images recorded using a multimode source fiber is substantially reduced.
A spatially-multiplexed swept-source optical coherence tomography
(SM-SS-OCT) system for rapid acquisition of B-scans of tissue microstructure is described, we believe, for the first time.
SM-SS-OCT instrumentation is similar to that of traditional Swept Source OCT (SS-OCT), which uses a widely tunable (~100 nm) laser source to obtain high-resolution images of biological tissue. However, SM-SS- OCT may be considered an improvement over SS-OCT in terms of efficient usage of the wide spectral bandwidth afforded by the frequency-tunable lasers in SS-OCT systems. Commercially available swept-source lasers regularly achieve extremely narrow line widths (~150 KHz), allowing for SS-OCT A-scan depths on the order of meters. Since imaging tissue to such depths is infeasible, the
meters-long depth ranging capability of SS-OCT may be utilized for spatially multiplexing many A-scans, each to lesser depth. We achieve this spatial multiplexing by rapidly scanning all lateral positions of the tissue repetitively while simultaneously scanning the laser wavelength continuously, and using appropriate signal processing to reconstruct a B-scan image from acquired data. Our fiber-based design lends itself towards use in endoscopic applications, and our results suggest that SM-SS-OCT can provide rapid acquisition of B-scans, with potential for depth-resolved visualization of transient processes in biological tissue.
Functional imaging of clear and tissue-simulating phantoms using phase-resolved swept-source spectroscopic OCT (PhS-SSOCT) is described. Superior sensitivity of PhS-SSOCT technique to monitor ultra-small changes in sample refractive index is demonstrated using aqueous solutions of glucose and aqueous suspensions of polystyrene microspheres and glucose. Glucose-induced changes in the phase are found to be 0.039 rad/mM and 0.037 rad/mM in the 200 μm-thick cell for clear and turbid media, respectively, that is in good agreement with our previous data obtained using differential-phase time-domain OCT. Obtained results suggest that PhS-SSOCT has a potential for noninvasive, depth-resolved, real-time quantitative monitoring of concentrations of glucose and other analytes with high accuracy.
We report on work to construct a dispersion control system using a spatial light modulator (SLM) in conjunction with optical coherence tomography. To test feasibility of the dispersion control system, we simulate the group delay by water, analyze phase of coherence function, and compute the dispersion compensating function for the SLM. In addition, a unique interferometer configuration utilizing the SLM is described. We verify that dispersion information in optical coherence imaging can be measured by phase analysis of coherence function in optical frequency domain and modified for a variety of applications using the control system.
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