Fibroblasts are important players in proliferation, invasion, migration and metastasis of cancer cells. Until now, histological methods such as hematoxylin and eosin (H and E) staining are still a gold standard to assess fibroblasts in traditional histopathology. Multiphoton microscopy (MPM) has become a key technology for biological tissues imaging without staining at the cellular level. In this study, MPM based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was used to identify the fibroblasts in breast cancer tissue. Normal fibroblasts with spindle shape and abnormal fibroblasts with stellate shape could be clearly obtained by MPM.
Nicotinamide adenine dinucleotide (NADH) and Flavin adenine dinucleotide (FAD) are the endogenous fluorophores in cells. Optical redox ratio determined by the ratio of NADH and FAD was an important oxygenic indicator to reflect cell metabolism. Heat shock protein 70 (HSP70) has been recognized as a target for antitumor therapy. In this study, the optical redox ratio was used to evaluate the metabolic changes after the synergistic treatment of tamoxifen and HSP70 inhibitor in MCF-7 breast cancer cells based on optical metabolic imaging. In addition, the cell viability was also measured after the treatments of tamoxifen and HSP70 inhibitor by MTT assay. We observed that changes of the redox ratio and the cell viability with synergistic treatment of tamoxifen and HSP70 inhibitor were more obvious than the results of the sole treatment of tamoxifen or HSP70 inhibitor. These results indicated that the inhibition of HSP70 improve drug response of MCF-7 breast cancer cells.
Resting heart rate (RHR) is considered an important biomedical indicator to evaluate cardiovascular function. High RHR is an important prognostic factor for sudden cardiac death and heart failure in the general population, and especially among patients with known cardiac disease. The imaging photoplethysmography (IPPG) technology is used to achieve the accurate detection of RHR signal, which has the advantages of low cost, simple operation, fast acquisition speed etc. In this paper, we propose a new simple, inexpensive and easy-to-use method to measure the RHR in vivo. The result shows that Fast Fourier Transform with Hamming window filters, band-pass filter gives more accurate results. The color change of the fingertip is enlarged by using the mobile phone camera. From the distribution of color change of the fingertip, the RHR is estimated with the primary calibration result of the relationship between color variation and the blood volume change.
Heat shock (HS) is one of the best-studied exogenous cellular stresses, and all cellular compartments and metabolic processes are involved in HS response. The heat shock proteins (Hsps) expression enhanced during HS mainly localized in subcellular compartments, such as cytosol, endoplasmic reticulum and mitochandria. The major inducible heat shock protein 70 (Hsp70) modulate cellular homeostasis and promote cellular survival by blocking a caspase independent cell death through its association with apoptosis inducing factor. Mitochondria as the critical elements of HS response that participate in key metabolic reactions, and the changes in mitochonrial morphology may impact on mitochondrial metabolism. In this paper, the changes of mitorchondrial morphology in breast cancer cell have been monitored in real time after heat shock (43 ℃ ) by the fluorescence imaging, and the influence of Hsp70 inhibitor on mitochandrial structures have also been investigated. Then the information of mitochondrial metabolism which can be characterized by the level of the mitochondrial membrane potential has also been obtained wihout/with the treatment of Hsp70 inhibitor. Our data indicated that the mitochandrial morphology were related with the mitochandrial membrane potential, and the mitochandrial membrane potential was influenced significantly with the treatment of Hsp70 inhibitor during HS.
Two sets of in vivo female cheek skin epidermis images were analyzed through gray level co-occurrence matrix (GLCM) and fast fourier transform (FFT). One set was derived from women in their 20s and the other from women more than 60 years of age. GLCM was used to evaluate the texture features of the regions of interest within the cheek epidermis, and texture classification was subsequently performed. During texture classification, 25 images (320×240 pixels) in each age set were randomly selected. Three texture features, i.e., energy, contrast, and correlation, were obtained from the skin images and analyzed at four orientations (0°, 45°,90°, and 135°), accompanied by different distances between two pixels. The textures of the different aging skins were characterized by FFT, which provides the dermatoglyph orientation index. The differences in the textures between the young and old skin samples can be well described by the FFT dermatoglyph orientation index. The texture features varied among the different aging skins, which provide a versatile platform for differentiating the statuses of aging skins.
The 632nm wavelength low intensity He-Ne laser was used to irradiated on 15 mice which had skin wound. The
dynamic changes and wound healing processes were observed with nonlinear spectral imaging technology. We observed
that:(1)The wound healing process was accelerated by the low-level laser therapy(LLLT);(2)The new tissues produced
second harmonic generation (SHG) signals. Collagen content and microstructure differed dramatically at different time
pointed along the wound healing. Our observation shows that the low intensity He-Ne laser irradiation can accelerate the
healing process of skin wound in mice, and SHG imaging technique can be used to observe wound healing process,
which is useful for quantitative characterization of wound status during wound healing process.
The diffuse light detected from tissue surface contains the information about the tissue optical parameters. How
to extract tissue optical parameters from the reflectance light with accuracy plays a significant role in optical
diagnosis. A nonlinear least square fitting algorithm was used to display the fitting accuracy influenced by the
fitting start time, the fitting end time, the two parameters fitting ( μa , μ's ) and the three parameters fitting ( μa , μ's , amplitude factor A ) on the basis of the MC simulation. A comparison is made between the results that to
use the diffuse reflectance data with a certain detecting angle to fit to the diffuse reflectance formula based on
the full angle detecting and to fit to the diffusion formula based on corresponding detecting angle respectively.
The influences of different nonlinear least square fitting algorithm on the fitting accuracy and on the fitting time
are also discussed. The result shows that the optical parameters determination is influenced by many factors. It
may play a significant role in improving the fitting accuracy of tissue optical parameters and in the tissue
noninvasive diagnosis.
Because of frequent exposure to carcinogens, the bronchus is prone to early pathologic alterations. The assessment of these early changes is of key significance in physiological studies and disease diagnosis of the bronchus. We utilize nonlinear optical microscopy (NLOM) to image mouse bronchial tissue based on intrinsic nonlinear optical contrast. Our results show that NLOM is effective for imaging the bronchial intact microstructural components, providing quantitative information about the biomorphology and biochemistry of tissue. Our findings also display that NLOM can provide a two-photon ratiometric redox fluorometry, based on mitochondrial signals and reduced pyridine nucleotide (NADH and NADPH) and oxidized flavoproteins (Fp) signals, to assess the metabolic state of the epithelial cells and chondrocytes. It was found that NLOM can offer a sensitive tool, based on the second-harmonic signal depth-dependent decay, to obtain quantitative information on the optical property of the stroma associated with normal and diseased tissue states. Our results suggest that with the advent of the clinical portability of typical nonlinear optical endoscopy, the NLOM technique has the potential to be applied in vivo to the clinical diagnosis and monitoring of bronchial disease.
A Monte Carlo algorithm to simulate the propagation of polarized light was used and the backscattered Mueller matrices
were calculated for various scattering and absorption coefficients and layered structures. Results show that scattering
produce a spatial re-distribution of light, while absorption only causes a uniform decrease in intensity. The major
contribution to backscattered Mueller matrix comes from weakly scattered photons. As scattering enhances the majority
of these photons exit from around the centre. result in image intensity increase around the centre and decrease at the edge.
For absorption strengthen only uniform intensity weakening was observed. Both can be easily differentiated by Mueller
matrix distribution patterns. A two-layer niodel with thin surface layer was also calculated, Mueller matrix represent
disparate alternant rule for upper and lower layer, which suggest possibilities for discriminating subtle epidennal
structure. All these results together indicate that discriminate optical properties in turbid media using backscattered
Mueller matrix is feasible.
Proc. SPIE. 6085, Complex Dynamics and Fluctuations in Biomedical Photonics III
KEYWORDS: Optical properties, Tissues, Diffusion, Reflectivity, Monte Carlo methods, Transmittance, Diffuse optical spectroscopy, Time resolved spectroscopy, Tissue optics, Algorithm development
The technology of time-resolved diffuse optical spectroscopy to determine optical parameters of biological tissues is reviewed in this paper. The theoretical model and some the development conditions are also presented. As an example, a Monte Carlo algorithm was developed to a two-layered tissue. Based on the numerical simulation, the structure information including the optical properties of the tissue can be obtained. In addition, the prospect of time-resolved diffuse optical spectroscopy technology is commented based on our opinion.
Proc. SPIE. 6084, Optical Interactions with Tissue and Cells XVII
KEYWORDS: Refractive index, Injuries, Skin, Microscopy, Collagen, Biopsy, Digital image correlation, In vivo imaging, Tissue optics, Animal model studies
Living cells and transparent specimen can be observed by the Differential Interference Contrast (DIC) microscopy. In this paper, the morphologic changes of mouse skin in vivo treated in several configurations by intense pulse light (IPL) were observed by a DIC microscopy. The differences of the images before and after the IPL irradiation were obtained and the mechanism was analyzed. In addition, the collagen recombination after the irradiation of IPL in skin was acquired.
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