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Optical coherence tomography is widely used in the non-invasive detection of skin cancer. We are proposing an image guiding system integrated in a full-field OCT (FF-OCT) system and an image matching algorithm to register the guiding image onto a dermoscopic image. This guiding system takes the objective optics of FF-OCT system to observe the same vision with OCT. The FOV of the guiding image and FF-OCT is 2*2mm2 and 500*500μm2 respectively. The spatial relation between OCT and the guiding image system is fixed, the OCT imaging location can also be identified. The guiding image is then matched into the clinically used dermoscopic image which has a 20*20 mm2 FOV by an image matching algorithm. The FF-OCT image can be registered on the dermoscopic image through the guiding image. This allows physicians to target the scanning area precisely in the lesion, and record the scanned point to confirm the full lesion. This invention improves the efficiency of the entire examination and allows to follow-ups the lesion at different time.

Mirau-type full-filed optical coherent tomography (FF-OCT) can provide imaging with cellular resolution section of human skin in vivo. In our previous work, the concept of dual-mode FF-OCT for cross-sectional (B-scan) and en face (E-scan) imaging with symmetrical illumination has been reported. This work presents a prototype of the dual-mode FF-OCT system and demonstrates the improvement of the efficiency for clinical applications. For clinical diagnosis, it is helpful to differentiate different types of nevus. Thus, an imaging tool capable of identifying the types of nevus is crucial. Our system can switch between two modes of imaging automatically by just clicking a button. The feature allows clinicians to explore the nevus more efficiently. While the B-scan images reveal the distribution of melanosomes, users can set a specific depth of the E-scan images to explore the morphology of surrounding skin cells instantly. The different types of nevus including junction nevus, intradermal nevus and compound nevus can be identified by using this dual-mode FF-OCT system.

Dynamic-focusing optical coherence tomography (OCT) can provide ~1-μm isotropic resolution in extended depth of field, and Mirau design is preferred because of lower weight. In previous works, we reported a Mirau-type OCT based on high-radiance spatially-incoherent source. However, the illumination intensity is limited by the etendue. Besides, coherent artifacts increases with the partially coherent illumination scheme. In this report, we introduce a method to narrow illumination linewidth to 5 μm and reduce coherence area to 1 μm2. With spatially incoherent illumination, optimized throughput and low-weight design, low-speckle B-scan can be acquired within 0.25 second.

A crystalline-fiber-based Mirau-type full-field optical coherence tomography (FF-OCT) system utilizing two partially coherent illumination modes is presented. Using a diode-pumped Ti:sapphire crystalline fiber with a high numerical aperture, spatially-incoherent broadband emission can be generated with high radiance. With two modes of different spatial coherence settings, either deeper penetration depth or higher B-scan rate can be achieved. In a wide-field illumination mode, the system functions like FF-OCT with partially coherent illumination to improve the penetration depth. In a strip-field illumination mode, a compressed field is generated on the sample, and a low-speckle B-scan can be acquired by compounding pixel lines within.

Significance: Melanocytic nevi represent the most common dermal melanocytic lesions in humans. Nevus is typically diagnosed clinically with the naked eye or with dermoscopy. However, it is essential to identify the type of nevus by invasive biopsy for histopathological examination. The use of noninvasive imaging tools can be used to evaluate the types of nevi to reduce unnecessary excisions of benign entities. Aim: To evaluate the feasibility of using en face and cross-sectional full-field optical coherence tomography (FF-OCT) in differentiation of melanocytic nevi that can facilitate the reduction of unnecessary excisions of benign entities. Approach: Dual-mode Mirau-type FF-OCT for cross-sectional imaging (B-scan) and en face imaging were used to distinguish the types of nevi. Results: Although the B-scan reveals the distribution of melanosomes, users can set a specific depth of the en face image to explore the morphology of surrounding skin cells instantly. According to the locations of nevus nests, the different types of nevi, including junction nevus and compound nevus, can be identified using this dual-mode FF-OCT system. Conclusions: Combining B-scan and en face imaging in vivo FF-OCT enables the examination and navigation of skin tissues in real time and in three dimensions.

Optical coherence microscopy (OCM) is an efficient tool to acquire cellular-resolution morphological images. In our previous study, we presented a Mirau-type OCM which provides both in vivo cross-sectional (B-scan) and en face imaging with a broadband extended source. However, two issues were encountered in B-scan mode. One is the illumination efficiency is low because the illumination area is much larger than the area of interest. The other is the image quality is degraded by linear artifacts. In this report, we use a symmetrical illumination design to improve the illumination efficiency and image quality of B-scan at the same time.

The encapsulation and/or surface modification can stabilize and protect the phosphorescence bio‐probes but impede their intravenous delivery across biological barriers. Here, a new class of biocompatible rhenium (ReI) diimine carbonyl complexes is developed, which can efficaciously permeate normal vessel walls and then functionalize the extravascular collagen matrixes as in situ oxygen sensor. Without protective agents, ReI‐diimine complex already exhibits excellent emission yield (34%, λ em = 583 nm) and large two‐photon absorption cross‐sections (σ 2 = 300 GM @ 800 nm) in water (pH 7.4). After extravasation, remarkably, the collagen‐bound probes further enhanced their excitation efficiency by increasing the deoxygenated lifetime from 4.0 to 7.5 µs, paving a way to visualize tumor hypoxia and tissue ischemia in vivo. The post‐extravasation functionalization of extracellular matrixes demonstrates a new methodology for biomaterial‐empowered phosphorescence sensing and imaging., Biocompatible rhenium (ReI) diimine carbonyl complexes functionalize the extravascular collagen matrixes as in situ oxygen sensor. After tail‐vein delivery and vessel penetration, the probe can bind extravascular collagens and achieve stabilization enhanced two‐photon phosphorescence yields. Increased deoxygenated phosphorescence lifetime from 4.0 to 7.5 µs paves a way to visualize tumor hypoxia and tissue ischemia in vivo.

A single nanomaterial with multiple imaging contrasts and functions is highly desired for multiscale theragnosis. Herein, we demonstrate single 1-1.9 μm infrared-active FePt alloy nanoparticles (FePt NPs) offering unprecedented four-contrast-in-one molecular imaging - computed tomography (CT), magnetic resonance imaging (MRI), photoacoustic (PA) imaging, and high-order multiphoton luminescence (HOMPL) microscopy. The PA response of FePt NPs outperforms that of infrared-active gold nanorods by 3- to 5.6-fold under identical excitation fluence and particle concentrations. HOMPL (680 nm) of an isolated FePt NP renders spatial full-width-at-half-maximum values of 432 nm and 300 nm beyond the optical diffraction limit for 1230-nm and 920-nm excitation, respectively. The in vivo targeting function was successfully visualized using HOMPL, PA imaging, CT, and MRI, thereby validating FePt as a single nanomaterial system covering up to four types (Optical/PA/CT/MRI) of molecular imaging contrast, ranging from the microscopic level to whole-body scale investigation.

On the basis of an infrared femtosecond Cr:forsterite laser, we developed a semiquantitative method to analyze the microscopic distribution of bilirubins. Using 1230 nm femtosecond pulses, we selectively excited the two-photon red fluorescence of bilirubin dimers around 660 nm. Autofluorescences from other endogenous fluorophores were greatly suppressed. Using this distinct fluorescence measure, we found that poorly differentiated hepatocellular carcinoma (HCC) tissues on average showed 3.7 times lower concentration of bilirubins than the corresponding nontumor parts. The corresponding fluorescence lifetime measurements indicated that HCC tissues exhibited a longer lifetime (500 ps) than that of nontumor parts (300 ps). Similarly, oral cancer cell lines had longer lifetimes (>330 ps) than those of nontumor ones (250 ps). We anticipate the developed methods of bilirubin molecular imaging to be useful in diagnosing cancers or studying the dynamics of bilirubin metabolisms in live cells.

Traditional biopsy procedures require invasive tissue removal from a living subject, followed by time-consuming and complicated processes, so noninvasive in vivo virtual biopsy, which possesses the ability to obtain exhaustive tissue images without removing tissues, is highly desired. Some sets of in vivo virtual biopsy images provided by healthy volunteers were processed by the proposed cell segmentation approach, which is based on the watershed-based approach and the concept of convergence index filter for automatic cell segmentation. Experimental results suggest that the proposed algorithm not only reveals high accuracy for cell segmentation but also has dramatic potential for noninvasive analysis of cell nuclear-to-cytoplasmic ratio (NC ratio), which is important in identifying or detecting early symptoms of diseases with abnormal NC ratios, such as skin cancers during clinical diagnosis via medical imaging analysis.

In vivo higher-harmonic generation microscopy (HGM) performed on healthy human oral mucosa not only provides images with a

Recent clinical studies on human skin indicated that in vivo multi-harmonic generation microscopy (HGM) can achieve sub-micron resolution for histopathological analysis with a high penetration depth and leave no energy or photodamages in the interacted tissues. It is thus highly desired to apply HGM for in vivo mucosa histopathological diagnosis. In this paper, the first in vivo optical virtual biopsy of human oral mucosa by using epi-HGM is demonstrated. We modified an upright microscope to rotate the angle of objective for in vivo observation. Our clinical study reveals the capability of HGM to in vivo image cell distributions in human oral mucosa, including epithelium and lamina propria with a high penetration depth greater than 280 μm and a high spatial resolution better than 500 nm. We also found that the third-harmonic-generation (THG) contrast on nucleus depends strongly on its thicknesses, in agreement with a numerical simulation. Besides, 4% acetic acid was found to be able to enhance the THG contrast of nucleus in oral mucosa, while such enhancement was found to decay due to the metabolic clearance of the contrast enhancer by the oral mucosa. Our clinical study indicated that, the combined epi-THG and epi-second-harmonic-generation (SHG) microscopy is a promising imaging tool for in vivo noninvasive optical virtual biopsy and disease diagnosis in human mucosa.

We use memory kernel to study the spontaneous emission of an atom in a photonic crystal with the isotropic band and anisotropic band. Our studies show that the long-time memory of the spontaneous emission in the photonic crystal induces a fractal phenomenon. Therefore, the fractional calculus is a natural mathematics to describe the fractal phenomenon. When the atomic transition frequency lies within the allowed band, using the fractional calculus we show that there is no multiple-valued problem and no fractionalized steady-state inversion encountered in the previous studies [J. Mod. Opt. 41, 353 (1994)]. On the other hand. We also consider an atom with finite size effect leading to multipole contributions and yield the memory kernel with the same form as a point-like atom. However, the memory kernel including finite-size effect has different coupling constant.

In vivo harmonic generation microscopy (HGM) has been applied successfully in healthy human skin and can achieve a submicron resolution, similar to histopathologic examination, even at a penetration depth up to 270 μm. This study aims to investigate the clinical applicability of HGM imaging for differential diagnosis of nonmelanoma pigmented skin lesions. A total of 42 pigmented skin tumors, including pigmented basal cell carcinoma, melanocytic nevus, and seborrheic keratosis were evaluated by HGM ex vivo or in vivo. Based on the standard histopathologic characteristics, we established the corresponding HGM imaging criteria for each pigmented tumor. Diagnostic performance of HGM for differentiating nonmelanoma pigmented skin tumors was evaluated through the observers’ direct general assessment (overall evaluation) or the presence of two imaging criteria with the highest sensitivity and specificity (major criteria evaluation). Our results show that, based on the direct general assessment, the sensitivity is 92% [95% confidence interval (CI): 67 to 97%] and the specificity is 96% (95% CI: 83 to 99%); by major criteria evaluation, 94% sensitivity (95% CI: 70 to 99%) and 100% specificity (95% CI: 87 to 100%) are achieved. Our study indicates that HGM serves as a promising histopathological examination tool for noninvasive differential diagnostics of nonmelanoma pigmented skin tumors.

A collagen second harmonic generation (SHG) and third harmonic generation (THG) image analysis algorithm based on feature extraction and classification for diabetes disease determination is developed in the paper. It is designed to find the early symptoms of diabetes by using the proposed method to calculate diabetes estimation indicator. The algorithm detects the basic diabetes featured elements on images of human dermis skin. The proposed algorithm consists of the processes of smoothing, adaptive three-region thresholding, binarization, segmentation, feature extraction, and diabetes disease determination. This feature classification criterion provides a new viewpoint to help diabetes diagnosis.

Morphology and distribution of melanocytes are critical imaging information for the diagnosis of melanocytic lesions. However, how to image intratumoral melanocytes noninvasively in pigmented skin tumors is seldom investigated. Third-harmonic generation (THG) is shown to be enhanced by melanin, whereas high accuracy has been demonstrated using THG microscopy for in vivo differential diagnosis of nonmelanocytic pigmented skin tumors. It is thus desirable to investigate if label-free THG microscopy was capable to in vivo identify intratumoral melanocytes. In this study, histopathological correlations of label-free THG images with the immunohistochemical images stained with human melanoma black (HMB)-45 and cluster of differentiation 1a (CD1a) were made. The correlation results indicated that the intratumoral THG-bright dendritic-cell-like signals were endogenously derived from melanocytes rather than Langerhans cells (LCs). The consistency between THG-bright dendritic-cell-like signals and HMB-45 melanocyte staining showed a kappa coefficient of 0.807, 84.6% sensitivity, and 95% specificity. In contrast, a kappa coefficient of −0.37, 21.7% sensitivity, and 30% specificity were noted between the THG-bright dendritic-cell-like signals and CD1a staining for LCs. Our study indicates the capability of noninvasive label-free THG microscopy to differentiate intratumoral melanocytes from LCs, which is not feasible in previous in vivo label-free clinical-imaging modalities.

Skin aging is an important issue in geriatric and cosmetic dermatology. To quantitatively analyze changes in keratinocytes related to intrinsic aging, we exploited a 1230 nm-based in vivo harmonic generation microscopy, combining second- and third-harmonic generation modalities. 52 individuals (21 men and 31 women, age range 19-79) were examined on the sun-protected volar forearm. Through quantitative analysis by the standard algorithm provided, we found that the cellular and nuclear size of basal keratinocytes, but not that of granular cells, was significantly increased with advancing age. The cellular and nuclear areas, which have an increase of 0.51 μm(2) and 0.15 μm(2) per year, respectively, can serve as scoring indices for intrinsic skin aging.

Traditional biopsy procedure requires invasive tissue removal from a living subject followed by time-consuming complicatedly processing, so noninvasive in vivo virtual biopsy is a highly desired technique which own ability to obtain exhaustive tissue images without removing tissues from subjects. Some sets of in vivo virtual biopsy images provided by some healthy volunteers are processed by our cell segmentation approach based on marker-controlled gradient watershed algorithm to isolate the nuclei and cytoplasm and also evaluate their Nuclear-to-Cytoplasmic (NC) ratio. From our experimental results, our algorithm has significant potential for in vivo cell segmentation and NC ratio analysis to identify or detect the early symptoms of some skin diseases with abnormal NC ratios, such as skin cancers in clinical diagnosis.

We suggest a better mathematical method, fractional calculus, for studying the behavior of the atom?field interaction in photonic crystals. By studying the spontaneous emission of an atom in a photonic crystal with a one-band isotropic model, we found that the long-time inducing memory of the spontaneous emission is a fractional phenomenon. This behavior could be well described by fractional calculus. The results show no steady photon?atom bound state for the atomic resonant transition frequency lying in the proximity of the allowed band edge which was encountered in a previous study (Woldeyohannes and John 2003?J. Opt. B: Quantum Semiclass. Opt.?5?R43). The correctness of this result is validated by the 'cut-off smoothing' density of photon states (DOS) with fractional calculus. By obtaining a rigorous solution without the multiple-valued problem for the system, we show that the method of fractional calculus has a logically concise property.

In this paper, we demonstrate that the third-harmonic-generation contrast on epithelial nuclei in vivo observed in human oral cavity depends strongly on the thicknesses, rather than nonlinear susceptibility, of cell nuclei.

Without cavity dumping or external amplification, we report a femtosecond Cr:forsterite laser with a 1.4 W output power and 2 W in continuous wave (CW) operated with a crystal temperature of 267 K. In the femtosecond regime, the oscillator generates Kerr-lens-mode-locked 84 fs pulses with a repetition rate of 85 MHz, corresponding to a high 16.5 nJ pulse energy directly from a single Cr:forsterite resonator. This intense femtosecond Cr:forsterite laser is ideal to pump varieties of high power fiber light sources and could be thus ideal for many biological and spectroscopy applications.

BACKGROUND: Atrial fibrosis creates a vulnerable tissue for atrial fibrillation (AF), but the spatial disarray of collagen fibers underlying atrial fibrosis is not fully elucidated. OBJECTIVE: This study hypothesizes that harmonics optical microscopy can illuminate the spatial mal-alignment of collagen fibers in AF via a layer-by-layer approach. PATIENTS AND METHODS: Atrial tissues taken from patients who underwent open-heart surgery were examined by harmonics optical microscopy. Using the two-dimensional Fourier transformation method, a spectral-energy description of image texture was constituted and its entropy was used to quantify the mal-alignment of collagen fibers. The amount of collagen fiber was derived from its area ratio to total atrial tissue in each image. Serum C-terminal pro-collagen pro-peptide (CICP), pro-matrix metalloproteinase-1 (pro-MMP-1), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were also evaluated. RESULTS: 46 patients were evaluated, including 20 with normal sinus rhythm and 26 with AF. The entropy of spectral-energy distribution of collagen alignment was significantly higher in AF than that in sinus rhythm (3.97 ± 0.33 vs. 2.80 ± 0.18, p

Oral cancer ranked number four in both cancer incident and mortality in Taiwanese male population. Early disease diagnosis and staging is essential for its clinical success. However, most patients were diagnosed in their late disease stage as ideal prescreening procedures are yet to be developed especially when dealing with a large surface of precancerous lesions. Therefore, how to detect and confirm the diagnosis of these early stage lesions are of significant clinical value. Harmonic generation process naturally occurred in biological molecules and requires no energy deposition to the target molecule. Thus harmonic generation microscopy (HGM) could potentially serve as a noninvasive tool for screening of human oral mucosal diseases. The in vivo optical biopsy of human oral cavity with HGM could be achieved with high spatial resolution to resolve dynamic physiological process in the oral mucosal tissue with equal or superior quality but devoid of complicated physical biopsy procedures. The second harmonic generation (SHG) provide significant image contrast for biomolecules with repetitive structures such as the collagen fibers in the lamina propria and the mitotic spindles in dividing cells. The cell morphology in the epithelial layer, blood vessels and blood cells flow through the capillaries can be revealed by third harmonic generation (THG) signals. Tissue transparent technology was used to increase the optical penetration of the tissue. In conclusion, this report demonstrates the first in vivo optical virtual biopsy of human oral mucosa using HGM and revealed a promising future for its clinical application for noninvasive in vivo diseases diagnosis.

Cartilage matrix is damaged in diseased states such as osteoarthritis, while adult articular cartilage does not have the capacity to repair structural damage. A least invasive mean to diagnose these diseased states of human articular cartilage with a high spatial resolution is thus highly desired. In this paper, we present our harmonic generation microscopic studies on the human articular cartilage samples. Without any staining, third and second harmonic generation can provide strong contrast in chondrocytes and collagen matrix, respectively. Our study indicates the high capability of harmonic generation microscopy for future articular cartilage disease diagnosis.

Myocardial fibrosis, a common sequela of cardiac hypertrophy, has been shown to be associated with arrhythmias in experimental models. Some research has indicated that myocardial fibrosis plays an important role in predisposing patients to atrial fibrillation. Second harmonic generation (SHG) is an optically nonlinear coherent process to image the collagen network. In this presentation, we observe the SHG images of the collagen matrix in atrial myocardium and we analyzed of collagen fibers arrangement by using Fourier-transform analysis. Moreover, comparing the SHG images of the collagen fibers in atrial myocardium between normal sinus rhythm (NSR) and atrial fibrillation (AF), our result indicated that it is possible to realize the relation between myocardial fibrosis and AF.

Combination of turbo codes with partial response (PR) channels was comprehensively studied in magnetic recording systems. In this paper, we investigate a decoding scheme called 'iterative tri-stage decoding' for the turbo code as the outer code and the precoded PR channel as the inner code. This decoding scheme reduces the computational complexities by reducing the number of iterations without degrading the coding gain.

Chronological skin aging is associated with flattening of the dermal-epidermal junction (DEJ), but to date no quantitative analysis focusing on the aging changes in the dermal papillae (DP) has been performed. The aim of the study is to determine the architectural changes and the collagen density related to chronological aging in the dermal papilla zone (DPZ) by in vivo harmonic generation microscopy (HGM) with a sub-femtoliter spatial resolution. We recruited 48 Asian subjects and obtained in vivo images on the sun-protected volar forearm. Six parameters were defined to quantify 3D morphological changes of the DPZ, which we analyzed both manually and computationally to study their correlation with age. The depth of DPZ, the average height of isolated DP, and the 3D interdigitation index decreased with age, while DP number density, DP volume, and the collagen density in DP remained constant over time. In vivo high-resolution HGM technology has uncovered chronological aging-related variations in DP, and sheds light on real-time quantitative skin fragility assessment and disease diagnostics based on collagen density and morphology.

Third-harmonic generation (THG) microscopy has been reported to provide intrinsic contrast in elastic fibers, cytoplasmic membrane, nucleus, actin filaments, lipid bodies, hemoglobin, and melanin in human skin. For advanced molecular imaging, exogenous contrast agents are developed for a higher structural or molecular specificity. We demonstrate the potential of the commonly adopted tattoo dye as a THG contrast agent for in vivo optical biopsy of human skin. Spectroscopy and microscopy experiments were performed on cultured cells with tattoo dyes, in tattooed mouse skin, and in tattooed human skin to demonstrate the THG enhancement effect. Compared with other absorbing dyes or nanoparticles used as exogenous THG contrast agents, tattoo dyes are widely adopted in human skin so that future clinical biocompatibility evaluation is relatively achievable. Combined with the demonstrated THG enhancement effect, tattoo dyes show their promise for future clinical imaging applications.

Atrial fibrillation (AF) is the most common irregular heart rhythm and the mortality rate for patients with AF is approximately twice the mortality rate for patients with normal sinus rhythm (NSR). Some research has indicated that myocardial fibrosis plays an important role in predisposing patients to AF. Therefore, realizing the relationship between myocardial collagen fibrosis and AF is significant. Second-harmonic generation (SHG) is an optically nonlinear coherent process to image the collagen network. We perform SHG microscopic imaging of the collagen fibers in the human atrial myocardium. Utilizing the SHG images, we can identify the differences in morphology and the arrangement of collagen fibers between NSR and AF tissues. We also quantify the arrangement of the collagen fibers using Fourier transform images and calculating the values of angle entropy. We indicate that SHG imaging, a nondestructive and reproducible method to analyze the arrangement of collagen fibers, can provide explicit information about the relationship between myocardial fibrosis and AF.