Chlorinated hydrocarbons (CHCs) are among the most frequently detected volatile organic compounds in North American ground-water supplies. They have been widely used in the chemical industry as solvents and degreasing agents. The average concentration level for CHCs in our aquatic environment ranges from high-ppb for chloroform to low-ppb level for tetrachloroethylene(PCE). Methods used for detecting these compounds in the environment require high sensitivity and high selectivity. Conventional detection methods for CHCs include gas chromatography(GC) or GC/MS. Although these techniques do possess the required sensitivity and selectivity, they lack the capability of performing in-situ measurements.
Early detection of incipient caries would allow dentists to provide more effective measures to delay or to reverse caries’ progression at earlier stage. Such earlier intervention could lead to improved oral health for the patients and reduced burden to the health system. Previously, we have demonstrated that the combination of morphological and biochemical information furnished by optical coherence tomography (OCT) and polarized Raman spectroscopy (PRS), respectively, provided a unique tool for dental caries management. In this study we will report the first pre-clinical caries detection system that includes a hand-held probe with a size slightly larger than a tooth brush. This probe presents a novel platform combining both OCT and PRS optics in a very tight space ideal for clinical practice. OCT cross-sectional images of near-surface enamel morphology are obtained with miniaturized MEMS scanning device and are processed in real-time to identify culprit regions. These regions are sequentially analyzed with polarized Raman spectroscopy for further confirmation. PRS is performed using 830nm laser line and four detection channels in order to obtain polarized Raman spectroscopic data, i.e. depolarization ratio of the hydroxyapatite Raman band at ~960 cm-1. A detailed description of this hand-held caries detector and ex-vivo/in-vivo test results will be presented.
Wound management is a challenging and costly problem that is growing in importance as people are living longer. Instrumental methods are increasingly being relied upon to provide objective measures of wound assessment to help guide management. Technologies that employ near-infrared (NIR) light form a prominent contingent among the existing and emerging technologies. We review some of these technologies. Some are already established, such as indocyanine green fluorescence angiography, while we also speculate on others that have the potential to be clinically relevant to wound monitoring and assessment. These various NIR-based technologies address clinical wound management needs along the entire healing trajectory of a wound.
In this study we present a novel image analysis methodology to quantify and to classify morphological details in tissue collagen fibril organization and lipid deposition. Co-localized collagen (second harmonic, SHG) and lipid (coherent Raman, CARS) images of atherosclerotic artery walls were acquired by a supercontinuum-powered multi-modal nonlinear microscope. Textural features based on the first-order statistics (FOS) and gray level co-occurrence matrix (GLCM) parameters were extracted from the SHG and CARS images. Multi-group classifications based on support vector machine of SHG and CARS images were subsequently performed to investigate the potential of texture analysis in providing quantitative descriptors of structural and compositional changes during disease progression. Using a rabbit model, different collagen remodeling and lipid accumulation patterns in disease tissues can be successfully tracked using these image statistics, thus providing a robust foundation for classification. When the variation of the CARS image features were tracked against the age of the rabbit, it was noticed that older animals (advanced plaques) present a more complex necrotic core containing high-lipid extracellular structures with various shapes and distribution. With combined FOS and GLCM texture statistics, we achieved reliable classification of SHG and CARS images acquired from atherosclerotic arteries with >90% accuracy, sensitivity and specificity. The proposed image analysis methodology can also be applied in a wide range of applications to evaluate conditions involving collagen re-modeling and prominent lipid accumulation.
Luminal atherosclerosis imaging was demonstrated by multimodal femtosecond CARS
microscopy (MM-CARS). Using a myocardial infarction-prone rabbit model of
atherosclerosis, this study demonstrated the utility of multimodal CARS imaging in
determining atherosclerotic plaque burden through two types of image analysis procedures.
Firstly, multimodal CARS images were evaluated using a signal-intensity parameter based
on intensity changes derived from the multi-channel data (e.g. TPEF, SHG and CARS) to
classify plaque burden within the vessel. Secondly, the SHG images that mainly correspond
to collagen fibrils were evaluated using a texture analysis model based on the first-order
statistical (FOS) parameters of the image histogram. Correlation between FOS parameters of
collagen images with atherosclerosis plaque burden was established. A preliminary study
of using spectroscopic CARS in identifying the different lipid components within the plaque
was also discussed.
Label-free imaging of bulk arterial tissue is demonstrated using a multimodal nonlinear optical microscope based on a photonic crystal fiber and a single femtosecond oscillator operating at 800 nm. Colocalized imaging of extracellular elastin fibers, fibrillar collagen, and lipid-rich structures within aortic tissue obtained from atherosclerosis-prone myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbits is demonstrated through two-photon excited fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering, respectively. These images are shown to differentiate healthy arterial wall, early atherosclerotic lesions, and advanced plaques. Clear pathological changes are observed in the extracellular matrix of the arterial wall and correlated with progression of atherosclerotic disease as represented by the age of the WHHLMI rabbits.
In this study we compare the single-photon autofluorescence and multi-photon emission spectra obtained from the
luminal surface of healthy segments of artery with segments where there are early atherosclerotic lesions. Arterial tissue
was harvested from atherosclerosis-prone WHHL-MI rabbits (Watanabe heritable hyperlipidemic rabbit-myocardial
infarction), an animal model which mimics spontaneous myocardial infarction in humans. Single photon fluorescence
emission spectra of samples were acquired using a simple spectrofluorometer set-up with 400 nm excitation. Samples
were also investigated using a home built multi-photon microscope based on a Ti:sapphire femto-second oscillator. The
excitation wavelength was set at 800 nm with a ~100 femto-second pulse width. Epi-multi-photon spectroscopic signals
were collected through a fibre-optics coupled spectrometer. While the single-photon fluorescence spectra of
atherosclerotic lesions show minimal spectroscopic difference from those of healthy arterial tissue, the multi-photon
spectra collected from atherosclerotic lesions show marked changes in the relative intensity of two-photon excited
fluorescence (TPEF) and second-harmonic generation (SHG) signals when compared with those from healthy arterial
tissue. The observed sharp increase of the relative SHG signal intensity in a plaque is in agreement with the known
pathology of early lesions which have increased collagen content.
Nonlinear optical (NLO) microscopy provides a minimally invasive optical method for
fast molecular imaging at subcellular resolution with 3D sectioning capability in thick,
highly scattering biological tissues. In the current study, we demonstrate the imaging
of arterial tissue using a nonlinear optical microscope based on photonic crystal fiber
and a single femto-second oscillator operating at 800nm. This NLO microscope system
is capable of simultaneous imaging extracellular elastin/collagen structures and lipid
distribution within aortic tissue obtained from coronary atherosclerosis-prone WHHLMI
rabbits (Watanabe heritable hyperlipidemic rabbit-myocardial infarction) Clear
pathological differences in arterial lumen surface were observed between healthy
arterial tissue and atherosclerotic lesions through NLO imaging.
Nonlinear optical imaging technologies offer some intriguing medical diagnostic applications. Examples include fast
imaging of elastin and collagen distributions in diseased tissues using two-photon fluorescence (TPF) and second
harmonic generation (SHG), respectively. The 3D sectioning capabilities and biochemical specificity that enable fast
imaging in highly scattering biological media lie at the heart of the appeal of these nonlinear approaches for medical
applications. One of these promising nonlinear techniques relies on the resonance enhancement of the third order
nonlinear susceptibility by a vibrational mode of a molecule. Coherent Anti-Stokes Raman Scattering (CARS) can
provide similar vibrational information as a spontaneous Raman spectrum. The technique has been shown to be orders
of magnitude more sensitive than spontaneous Raman, with video rate imaging demonstrated recently. In this work,
we investigate the potential use of broadband CARS spectroscopy and CARS imaging for biochemical analysis of
arterial tissue. Biochemical imaging data from broadband CARS is compared with spontaneous Raman
microspectroscopy. The broadband CARS system comprised of a single femtosecond-laser is presented in detail.
Issues related to data analysis, the advantages and current limitations of the CARS technique in biodiagnostics are
discussed.
Incipient dental caries lesions appear as white spots on the tooth surface; however, accurate detection of early
approximal lesions is difficult due to limited sensitivity of dental radiography and other traditional diagnostic tools. A
new fibre-optic coupled spectroscopic method based on polarized Raman spectroscopy (P-RS) with near-IR laser
excitation is introduced which provides contrast for detecting and characterizing incipient caries. Changes in polarized
Raman spectra are observed in PO43- vibrations arising from hydroxyapatite of mineralized tooth tissue.
Demineralization-induced morphological/orientational alteration of enamel crystallites is believed to be responsible for
the reduction of Raman polarization anisotropy observed in the polarized Raman spectra of caries lesions. Supporting
evidence obtained by polarized Raman spectral imaging is presented. A specially designed fibre-optic coupled setup for
simultaneous measurement of parallel- and cross-polarized tooth Raman spectra is demonstrated in this study.
Early dental caries detection facilitates implementation of non-surgical methods for arresting caries progression and
promoting tooth remineralization. We present a method based on Raman spectroscopy with near-IR laser excitation to
provide biochemical contrast for detecting and characterizing incipient carious lesions found in extracted human teeth.
Changes in Raman spectra are observed in PO43- vibrations arising from hydroxyapatite of mineralized tooth tissue. Examination of various intensities of the PO43- ν2, ν3, ν4 vibrations showed consistent increased intensities in spectra of carious lesions compared to sound enamel. The spectral changes are attributed to demineralization-induced alterations of
enamel crystallite morphology and/or orientation. This hypothesis is supported by reduced Raman polarization
anisotropy derived from polarized Raman spectra of carious lesions. Polarized Raman spectral imaging of carious
lesions found on whole (i.e. un-sectioned) tooth samples will also be presented.
Atherosclerosis is traditionally viewed as a disease of uncontrolled plaque growth leading to arterial occlusion. More
recently, however, occlusion of the arterial lumen is being viewed as an acute event triggered by plaque rupture and
thrombosis. An atheromatous plaque becomes vulnerable to sudden activation and/or rupture when a constellation of
processes are activated by various trigger mechanisms. There is growing evidence that the vulnerability (i.e.
susceptibility to rupture) and thrombogenic nature of the plaque need to be taken into account in the planning and
treatment of the disease. X-ray fluoroscopy and intravascular ultrasound, the current clinical diagnostic tools are not
capable of the providing a complete histological picture of the plaque region.
Intravascular diagnostic imaging of coronary atherosclerotic plaques by optical means to assess plaque, patient risk and
assist in planning treatment strategies represents the future in angioplasty treatment by interventional cardiologists. The
techniques which will enable a clinically acceptable and reliable intravascular diagnostic platform are currently being
investigated and compared to the clinical standard of histology.
Currently, we are investigating the use of a number of optical and imaging techniques for biochemical analysis of
arterial tissue including Raman, near infrared and fluorescence spectroscopies. Biochemical imaging will provide
compositional information on collagen, elastin, lipid and thrombogenic by-products as well as gauging inflammation
and tissue remodeling activity levels. To complement the functional biochemical imaging, optical coherence
tomography will be provide structural morphological imaging. The synergistic combination of functional and structural
imagery will provide the interventional cardiologist with a complete clinical picture of the atherosclerotic plaque region.
The clinician can use this diagnostic information to plan a personalized treatment procedure based on the entire clinical
presentation.
The early approximal caries lesion in enamel is observed clinically as a white spot and is difficult to detect and/or
monitor with current methods available to dentists. New methods with high sensitivity and specificity are required to
enable improved early dental caries diagnosis. Using unpolarized Raman spectroscopy to examine unsectioned teeth,
peak intensity changes in the phosphate (PO43-) vibrations (ν2, ν3 and ν4) were observed between spectra of sound and
carious enamel. However, there is little change in the ν1 vibration with this approach. In contrast, when tooth sections
were examined by unpolarized Raman spectroscopy, marked changes in the ν1 peak at 959 cm-1 were noted between
healthy and carious enamel. These differences suggest that sampling orientation play a role in understanding the spectral
changes. Using polarized Raman spectroscopy to examine unsectioned samples, cross polarized measurements from
sound enamel exhibited significant reduction of the ν1 peak compared with parallel polarized measurements. A similar
reduction was observed with carious enamel, however, the reduction was not as prominent. By calculating the
depolarization ratio of the area under the ν1 peak, sound enamel can be clearly distinguished from demineralized
regions. The spectral changes observed are attributed to changes in the structure and/or orientation of the apatite crystals
as a result of the acid demineralization process.
Early dental caries detection will facilitate implementation of nonsurgical methods for arresting caries progression and promoting tooth remineralization. We present a method that combines optical coherence tomography (OCT) and Raman spectroscopy to provide morphological information and biochemical specificity for detecting and characterizing incipient carious lesions found in extracted human teeth. OCT imaging of tooth samples demonstrated increased light backscattering intensity at sites of carious lesions as compared to the sound enamel. The observed lesion depth on an OCT image was approximately 290 µm matching those previously documented for incipient caries. Using Raman microspectroscopy and fiber-optic-based Raman spectroscopy to characterize the caries further, spectral changes were observed in PO vibrations arising from hydroxyapatite of mineralized tooth tissue. Examination of various ratios of PO 2, 3, 4 vibrations against the 1 vibration showed consistent increases in carious lesions compared to sound enamel. The changes were attributed to demineralization-induced alterations of enamel crystallite morphology and/or orientation. OCT imaging is useful for screening carious sites and determining lesion depth, with Raman spectroscopy providing biochemical confirmation of caries. The combination has potential for development into a new fiber-optic diagnostic tool enabling dentists to identify early caries lesions with greater sensitivity and specificity.
Early dental caries result from destruction of the tooth's outer mineral matrix by acid-forming bacteria found in dental plaques. Early caries begin as surface disruptions where minerals are leached from the teeth resulting in regions of decreased mineral matrix integrity. Visually, these early carious regions appear as white spots due to the higher backscattering of incident light. With age these areas may become stained by organic compounds. Optical coherence tomography (OCT) examination of human teeth demonstrates a difference in penetration depth of the OCT signal into the carious region in comparison with sound enamel. However, while OCT demonstrates a structural difference in the enamel in the region of the caries, this technique provides little insight into the source of this difference. Raman spectroscopy provides biochemical measures derived from hydroxyapatite within the enamel as well as information on the crystallinity of the enamel matrix. The differences in the biochemical and morphological features of early caries and intact sound enamel are compared. Histological thin sections confirm the observations by OCT morphological imaging while Raman spectroscopy allows for biochemical identification of carious regions by a non-destructive method. Visual examination and conventional radiographic imaging of the intact tooth are used in clinical assessment prior to optical measurements. The combination of OCT, Raman spectroscopy and thin section histology aid in determining the changes that give rise to the visual white spot lesions.
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