The Experts below are selected from a list of 7323 Experts worldwide ranked by ideXlab platform
Michael J. Levene - One of the best experts on this subject based on the ideXlab platform.
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Multiphoton Microscopy with clearing for three dimensional histology of kidney biopsies.
Biomedical optics express, 2016Co-Authors: Eben Olson, Michael J. Levene, Richard TorresAbstract:We present a Multiphoton Microscopy approach with clearing optimized for pathology evaluation producing image quality comparable to traditional histology. Use of benzyl alcohol/benzyl benzoate with 4',6-diamidino-2-phenylindole and eosin in an optimized imaging setup results in optical sections nearly indistinguishable from traditionally-cut sections. Application to human renal tissue demonstrates diagnostic-level image quality can be maintained through 1 millimeter of tissue. Three dimensional perspectives reveal changes of glomerular capsule cells not evident on single sections. Collagen-derived second harmonic generation can be visualized through entire biopsies. Multiphoton Microscopy with clearing has potential for increasing the yield of histologic evaluation of biopsy specimens.
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Multiphoton Microscopy with Clearing for Renal Pathology
Biomedical Optics 2016, 2016Co-Authors: Eben Olson, Michael J. Levene, Richard TorresAbstract:We present a Multiphoton Microscopy approach with clearing optimized for pathology evaluation yielding quality comparable to traditional histology. Application to human renal tissue reveals pathologic findings not evident on single slide sections.
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Multiphoton Microscopy of cleared human tissue for 3D histology
Optics in the Life Sciences, 2013Co-Authors: Michael J. Levene, Kevin C. Zhou, Sam Vesuna, Richard TorresAbstract:Multiphoton Microscopy of optically cleared human biopsies can create a true 3D histopathology. We present recent advances in clearing protocols that reduce the total clearing time to
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Microprisms for Chronic In Vivo Multiphoton Microscopy of Cortex
Frontiers in Optics 2011 Laser Science XXVII, 2011Co-Authors: Michael J. LeveneAbstract:We demonstrate the use of microprisms for chronic in vivo Multiphoton Microscopy of mouse cortex. These prisms enable a point-of-view more typical of ex-vivo, cortical slice preparations, but in an in vivo context.
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Multiphoton Microscopy of cleared mouse organs
Journal of Biomedical Optics, 2010Co-Authors: Sonia Parra, Joseph P. Zinter, Thomas H. Chia, Michael J. LeveneAbstract:Typical imaging depths with Multiphoton Microscopy (MPM) are limited to less than 300 µm in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs with penetration depths and fields of view in excess of 2 mm. MPM enables the creation of large 3-D data sets with flexibility in pixel format and ready access to intrinsic fluorescence and second-harmonic generation. We present high-resolution images and 3-D image stacks of the brain, small intestine, large intestine, kidney, lung, and testicle with image sizes as large as 4096×4096 pixels.
Thomas H. Chia - One of the best experts on this subject based on the ideXlab platform.
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Multiphoton Microscopy of cleared mouse organs
Journal of Biomedical Optics, 2010Co-Authors: Sonia Parra, Joseph P. Zinter, Thomas H. Chia, Michael J. LeveneAbstract:Typical imaging depths with Multiphoton Microscopy (MPM) are limited to less than 300 µm in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs with penetration depths and fields of view in excess of 2 mm. MPM enables the creation of large 3-D data sets with flexibility in pixel format and ready access to intrinsic fluorescence and second-harmonic generation. We present high-resolution images and 3-D image stacks of the brain, small intestine, large intestine, kidney, lung, and testicle with image sizes as large as 4096×4096 pixels.
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Microprisms for In Vivo Multiphoton Microscopy of Mouse Cortex
Biophysical Journal, 2010Co-Authors: Michael J. Levene, Thomas H. ChiaAbstract:Fluorescence Microscopy of cortical slices, yielding ready access to all six layers of cortex, has proven to be a powerful technique in neurophysiology, however it lacks the context of in vivo experiments. In vivo Microscopy, primarily Multiphoton Microscopy, provides this context but without ready access to deeper layers and typically involves imaging of a field-of-view that is roughly parallel to the cortical layers. Needle-like gradient index (GRIN) lenses have been used as invasive relay lenses to access deeper brain structures, however these lenses damage the apical dendrites of the neurons of interest during insertion into the cortex, and are therefore of limited use for functional cortical imaging.We present here the use of micro-prisms for performing in vivo Multiphoton Microscopy of mouse cortex. Small (∼1 mm ) prisms with a reflective coating on the hypotenuse act as a miniature periscope, rotating the image plane from one parallel to the cortical layers to one that is perpendicular to the layers. This enables simultaneous imaging of the entire thickness of cortex, much as is done it cortical slice preparations, while maintaining a large degree of the in vivo context.
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Microprisms for in vivo Multiphoton Microscopy of Cortex
Advances in Imaging, 2009Co-Authors: Michael J. Levene, Thomas H. ChiaAbstract:We demonstrate the use of microprisms for in vivo Multiphoton Microscopy of mouse cortex. These prisms enable a point-of-view more typical of ex vivo, cortical slice preparations, but in an in vivo context.
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Microprisms for in vivo Multiphoton Microscopy of Mouse Cortex
Frontiers in Optics 2009 Laser Science XXV Fall 2009 OSA Optics & Photonics Technical Digest, 2009Co-Authors: Thomas H. Chia, Michael J. LeveneAbstract:Microprisms inserted into the cortex of mouse enable in vivo Multiphoton Microscopy, rotating the field-of-view from parallel to perpendicular to the surface of cortex and allowing imaging of the full cortical thickness.
Chen-yuan Dong - One of the best experts on this subject based on the ideXlab platform.
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Enhanced Multiphoton Microscopy of bilogical tissue with optical clearing
Clinical and Biomedical Spectroscopy and Imaging III, 2013Co-Authors: Vladimir A. Hovhannisyan, Ara Ghazaryan, Chang-seok Kim, Chen-yuan DongAbstract:Time-lapsed, three-dimensional Multiphoton Microscopy showed that application of air-drying and glycerol to animal tissue induced a well-expressed optical clearing. The effect was dynamic, reversible process, and can be used to enhance capabilities of nonlinear imaging.
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Multiphoton Microscopy imaging of developing tooth germs
Journal of the Formosan Medical Association = Taiwan yi zhi, 2012Co-Authors: Pei-yu Pan, Chen-yuan Dong, Wei-liang Chen, Rung-shu Chen, Chih-liang Ting, Min-huey ChenAbstract:Background/Purpose Traditionally, tooth germ is observed by histological investigation with hematoxylin and eosin stain and information may loss during the process. The purpose of this study is to use Multiphoton laser fluorescence Microscopy to observe the developing tooth germs of mice for building up the database of the images of tooth germs and compare with those from conventional histological analysis. Methods Tooth germs were isolated from embryonic and newborn mice with age of Embryonic Day 14.5 and Postnatal Days 1, 3, 5, and 7. Results Comparison of the images of tooth germ sections in Multiphoton Microscopy with the images of histology was performed for investigating the molar tooth germs. It was found that various signals arose from different structures of tooth germs. Pre-dentin and dentin have strong second-harmonic generation signals, while ameloblasts and enamel tissues were shown with strong autofluorescence signals. Conclusion In this study, a novel Multiphoton Microscopy database of images from developing tooth germs in mice was set up. We confirmed that Multiphoton laser Microscopy is a powerful tool for investigating the development of tooth germ and is worthy for further application in the study of tooth regeneration.
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Multiphoton Microscopy in dermatological imaging.
Journal of dermatological science, 2009Co-Authors: Tsung-hua Tsai, Chen-yuan Dong, Shiou-hwa Jee, Sung-jan LinAbstract:A minimally invasive imaging modality that provides both cellular and extracellular structural information with subcellular resolution is helpful for clinical diagnosis as well as basic laboratory research in dermatology. Multiphoton Microscopy (MPM), using femtosecond laser as the light source, is efficient in non-linear excitation of endogenous fluorophores and induction of second harmonic generation signals from non-centrosymmetric biomolecules such as collagen. This imaging modality is minimally invasive in the sense that much of the traditional histological procedures can be bypassed en route to obtain morphological and structural information of high scattering skin tissues. This unique feature has allowed clinical dermatological diagnosis, both ex vivo and in vivo. In addition to discussing the basic principles of Multiphoton Microscopy, this review is aimed at emphasizing its specific applications to dermatological imaging, including characterizing stratum corneum structures, visualizing and quantifying transcutaneous drug delivery, detecting skin cancers, exploring collagen structural transitions, and monitoring laser-skin interactions.
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Visualizing laser-skin interaction in vivo by Multiphoton Microscopy
Journal of biomedical optics, 2009Co-Authors: Tsung-hua Tsai, Shiou-hwa Jee, Chen-yuan Dong, Jung Yi Chan, Jin Ning Lee, Woan Ruoh Lee, Sung-jan LinAbstract:Recently, Multiphoton Microscopy has gained much popularity as a noninvasive imaging modality in biomedical research. We evaluate the potential of Multiphoton Microscopy for monitoring laser-skin reaction in vivo. Nude mouse skin is irradiated with an erbium:YAG laser at various fluences and immediately imaged by a Multiphoton microscope. The alterations of cutaneous nonlinear optical properties including Multiphoton autofluorescence and second-harmonic generation associated with laser irradiation are evaluated morphologically and quantitatively. Our results show that an erbium:YAG laser at a low fluence can selectively disrupt the stratum corneum, and this alteration may account for the penetration enhancing effect of laser-assisted transcutaneous drug delivery. At a higher fluence, the zone of tissue ablation as well as the disruption of the surrounding stratum corneum, keratinocytes, and dermal extracellular matrix can be better characterized by Multiphoton Microscopy as compared with conventional histology. Furthermore, the degree of collagen damage in the residual thermal zone can be quantified by second-harmonic generation signals, which have significant difference between control skin, skin irradiated with a 1.5-, 8-, and 16-J/cm2 erbium:YAG laser (P
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Imaging tissue engineering scaffolds using Multiphoton Microscopy.
Microscopy research and technique, 2008Co-Authors: Yen Sun, Sung-jan Lin, Shiou-hwa Jee, Hsin-yuan Tan, Hsuan-shu Lee, Tzu-yu Lin, Tai-horng Young, Wei-liang Chen, Chen-yuan DongAbstract:In this study, we combined two-photon autofluorescence and second harmonic generation imaging to investigate the three-dimensional microstructure and nonlinear optical properties of tissue engineering scaffolds. We focused on five different types of scaffold materials commonly used in tissue engineering, including: open-cell polylactic acid, polyglycolic acid, collagen composite scaffold, collagraft bone graft matrix strip, and nylon. By the use of Multiphoton Microscopy and a motorized stage, we obtained high resolution, spectrally resolved structural information of the scaffolds over large areas or in three-dimensions. Our results show that the nonlinear optical properties of the scaffolds will enable us to spectrally and morphologically distinguish the different types of scaffold materials investigated. We envision Multiphoton Microscopy to be a useful technique in tissue engineering applications in understanding the interplay between cultured cells and the scaffold materials. Microsc. Res. Tech., 2008. © 2007 Wiley-Liss, Inc.
Sung-jan Lin - One of the best experts on this subject based on the ideXlab platform.
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Multiphoton Microscopy in dermatological imaging.
Journal of dermatological science, 2009Co-Authors: Tsung-hua Tsai, Chen-yuan Dong, Shiou-hwa Jee, Sung-jan LinAbstract:A minimally invasive imaging modality that provides both cellular and extracellular structural information with subcellular resolution is helpful for clinical diagnosis as well as basic laboratory research in dermatology. Multiphoton Microscopy (MPM), using femtosecond laser as the light source, is efficient in non-linear excitation of endogenous fluorophores and induction of second harmonic generation signals from non-centrosymmetric biomolecules such as collagen. This imaging modality is minimally invasive in the sense that much of the traditional histological procedures can be bypassed en route to obtain morphological and structural information of high scattering skin tissues. This unique feature has allowed clinical dermatological diagnosis, both ex vivo and in vivo. In addition to discussing the basic principles of Multiphoton Microscopy, this review is aimed at emphasizing its specific applications to dermatological imaging, including characterizing stratum corneum structures, visualizing and quantifying transcutaneous drug delivery, detecting skin cancers, exploring collagen structural transitions, and monitoring laser-skin interactions.
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Visualizing laser-skin interaction in vivo by Multiphoton Microscopy
Journal of biomedical optics, 2009Co-Authors: Tsung-hua Tsai, Shiou-hwa Jee, Chen-yuan Dong, Jung Yi Chan, Jin Ning Lee, Woan Ruoh Lee, Sung-jan LinAbstract:Recently, Multiphoton Microscopy has gained much popularity as a noninvasive imaging modality in biomedical research. We evaluate the potential of Multiphoton Microscopy for monitoring laser-skin reaction in vivo. Nude mouse skin is irradiated with an erbium:YAG laser at various fluences and immediately imaged by a Multiphoton microscope. The alterations of cutaneous nonlinear optical properties including Multiphoton autofluorescence and second-harmonic generation associated with laser irradiation are evaluated morphologically and quantitatively. Our results show that an erbium:YAG laser at a low fluence can selectively disrupt the stratum corneum, and this alteration may account for the penetration enhancing effect of laser-assisted transcutaneous drug delivery. At a higher fluence, the zone of tissue ablation as well as the disruption of the surrounding stratum corneum, keratinocytes, and dermal extracellular matrix can be better characterized by Multiphoton Microscopy as compared with conventional histology. Furthermore, the degree of collagen damage in the residual thermal zone can be quantified by second-harmonic generation signals, which have significant difference between control skin, skin irradiated with a 1.5-, 8-, and 16-J/cm2 erbium:YAG laser (P
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Imaging tissue engineering scaffolds using Multiphoton Microscopy.
Microscopy research and technique, 2008Co-Authors: Yen Sun, Sung-jan Lin, Shiou-hwa Jee, Hsin-yuan Tan, Hsuan-shu Lee, Tzu-yu Lin, Tai-horng Young, Wei-liang Chen, Chen-yuan DongAbstract:In this study, we combined two-photon autofluorescence and second harmonic generation imaging to investigate the three-dimensional microstructure and nonlinear optical properties of tissue engineering scaffolds. We focused on five different types of scaffold materials commonly used in tissue engineering, including: open-cell polylactic acid, polyglycolic acid, collagen composite scaffold, collagraft bone graft matrix strip, and nylon. By the use of Multiphoton Microscopy and a motorized stage, we obtained high resolution, spectrally resolved structural information of the scaffolds over large areas or in three-dimensions. Our results show that the nonlinear optical properties of the scaffolds will enable us to spectrally and morphologically distinguish the different types of scaffold materials investigated. We envision Multiphoton Microscopy to be a useful technique in tissue engineering applications in understanding the interplay between cultured cells and the scaffold materials. Microsc. Res. Tech., 2008. © 2007 Wiley-Liss, Inc.
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Multiphoton Microscopy: a new paradigm in dermatological imaging
European journal of dermatology : EJD, 2007Co-Authors: Sung-jan Lin, Shiou-hwa Jee, Chen-yuan DongAbstract:In recent years, the non-linear optical imaging technique of Multiphoton Microscopy has gained significant popularity in biomedical imaging. Since optical imaging can provide detailed morphological information of biological structures, Multiphoton Microscopy holds great promise as a potential clinical diagnostic tool of dermatological conditions. In this review, we will begin by discussing the basic principles of Multiphoton Microscopy, including the process of fluorescence and second harmonic generation. In addition, we will present the dermatological applications of Multiphoton imaging, including the diagnosis of basal cell carcinoma and the evaluation of skin photoaging. We also describe applications of this technique to transcutaneous drug delivery, melanoma imaging, skin diseases associated with extracellular matrix alterations and cutaneous microvascular observation. Finally, we will discuss the additional issues that need to be resolved before Multiphoton imaging can become a major diagnostic tool in clinical dermatology.
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Three-dimensional skin imaging using the combination of reflected confocal and Multiphoton Microscopy
Photonic Therapeutics and Diagnostics III, 2007Co-Authors: Ming-gu Lin, Sung-jan Lin, Shiou-hwa Jee, Hsin-yuan Tan, Tsung-hua Tsai, Wei-liang Chen, Chen-yuan DongAbstract:Reflected confocal Microscopy has been widely used in clinical application in dermatology. In recent years, Multiphoton Microscopy has also emerged as an important minimally invasive bioimaging technique for the skin. In this study, we combine reflected confocal Microscopy and Multiphoton Microscopy for skin imaging. In the epidermis, reflected confocal signals are expected to help in delineating cell borders while Multiphoton signals provide cytoplasmic morphologies. In the dermis, second harmonic generation signals provide the morphology of collagen fibers. When three-dimensional images are projected, the detailed distribution of cellular component and extracellular matrix in skin can be obtained. Properly developed, this technique is of great potential for in vivo clinical application.
Watt W. Webb - One of the best experts on this subject based on the ideXlab platform.
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Multiphoton Microscopy of Tissues for Medical Diagnostics
Frontiers in Optics 2011 Laser Science XXVII, 2011Co-Authors: Watt W. WebbAbstract:Multiphoton Microscopy for spectral imaging of tissue fluorescence at 500nm lateral resolution can provide early medical diagnostics for lung cancers, inflammation and infection with the potential to reduce the ubiquitous mortality associated with lung cancer. Article not available.
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Lens Design for Small Diameter in Vivo Endoscopic Multiphoton Microscopy
Biophysical Journal, 2011Co-Authors: Christopher M. Brown, Watt W. WebbAbstract:Multiphoton Microscopy has demonstrated the capacity to image tissue anatomy and morphology with high resolution optical depth sectioning and the potential for diagnosis of tissue health. We are building devices to transition Multiphoton imaging and diagnostic techniques from the laboratory to the clinic and hospital using in vivo Multiphoton endoscopy. In this instrument, lens design for a miniature (< 5 mm outer diameter) endoscopic Multiphoton Microscopy system presents a significant challenge. Lens system design goals include: wide field of view and moderate resolution image acquisition, high epifluorescent collection efficiency from scattering media, design for use with scanned optical fiber imaging systems, small device size and low cost manufacture. We present results for a three-successive element lens design that meets these criteria.
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Medical Endoscopes for Multiphoton Microscopy
Biophysical Journal, 2009Co-Authors: Hyungsik Lim, Watt W. WebbAbstract:We present a new design of medical endoscope, which is ideal for Multiphoton Microscopy of human tissue in vivo. We discuss the properties of our reflective objective lens and the advantages in deep tissue imaging.
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Multiphoton Microscopy in biological research
Current opinion in chemical biology, 2001Co-Authors: Rebecca M. Williams, Warren R. Zipfel, Watt W. WebbAbstract:From its conception a decade ago, Multiphoton Microscopy has evolved from a photonic novelty to an indispensable tool for gleaning information from subcellular events within organized tissue environments. Its relatively deep optical penetration has recently been exploited for subcellularly resolved investigations of disease models in living transgenic mice. Its enhanced spectral accessibility enables aberration-free imaging of fluorescent molecules absorbing in deep-UV energy regimes with simultaneous imaging of species having extremely diverse emission spectra. Although excited fluorescence is the primary signal for Multiphoton Microscopy, harmonic generation by Multiphoton scattering processes are also valuable for imaging species with large anharmonic modes, such as collagen structures and membrane potential sensing dyes.
