Anouk L. Post, Xu U Zhang, Nienke Bosschaart, T. V. van Leeuwen, H. Sterenborg, D. Faber
Both Optical Coherence Tomography (OCT) and Single Fiber Reflectance Spectroscopy (SFR) are used to determine various optical properties of tissue. We developed a method combining these two techniques to measure the scattering anisotropy (g1) and γ (=1-g2/1-g1), related to the 1st and 2nd order moments of the phase function. The phase function is intimately associated with the cellular organization and ultrastructure of tissue, physical parameters that may change during disease onset and progression. Quantification of these parameters may therefore allow for improved non-invasive, in vivo discrimination between healthy and diseased tissue. With SFR the reduced scattering coefficient and γ can be extracted from the reflectance spectrum (Kanick et al., Biomedical Optics Express 2(6), 2011). With OCT the scattering coefficient can be extracted from the signal as a function of depth (Faber et al., Optics Express 12(19), 2004). Consequently, by combining SFR and OCT measurements at the same wavelengths, the scattering anisotropy (g) can be resolved using µs’= µs*(1-g). We performed measurements on a suspension of silica spheres as a proof of principle. The SFR model for the reflectance as a function of the reduced scattering coefficient and γ is based on semi-empirical modelling. These models feature Monte-Carlo (MC) based model constants. The validity of these constants - and thus the accuracy of the estimated parameters - depends on the phase function employed in the MC simulations. Since the phase function is not known when measuring in tissue, we will investigate the influence of assuming an incorrect phase function on the accuracy of the derived parameters.
光学相干层析成像(OCT)和单光纤反射光谱(SFR)都用于确定组织的各种光学特性。我们开发了一种结合这两种技术的方法来测量与相函数的一阶和二阶矩相关的散射各向异性(g1)和γ (=1-g2/1-g1)。相功能与细胞组织和组织超微结构密切相关，在疾病发生和发展过程中可能发生变化的物理参数。因此，这些参数的量化可以改善健康和病变组织之间的非侵入性体内区分。利用SFR可以从反射光谱中提取散射系数和γ (Kanick et al.， Biomedical Optics Express 2(6)， 2011)。利用OCT可以从信号中提取出作为深度函数的散射系数(Faber et al.， Optics Express 12(19)， 2004)。因此，通过结合SFR和OCT在相同波长下的测量，散射各向异性(g)可以使用µs ' =µs*(1-g)来解决。我们对二氧化硅球悬浮液进行了测量，作为原理的证明。反射率的SFR模型是基于半经验模型的散射系数和γ的函数。这些模型具有基于蒙特卡罗(MC)的模型常数。这些常数的有效性以及估计参数的准确性取决于MC模拟中采用的相函数。由于在组织中测量时相函数是未知的，我们将研究假设不正确的相函数对导出参数精度的影响。
{"title":"Measuring the reduced scattering coefficient and γ with SFR spectroscopy: studying the phase function dependence (Conference Presentation)","authors":"Anouk L. Post, Xu U Zhang, Nienke Bosschaart, T. V. van Leeuwen, H. Sterenborg, D. Faber","doi":"10.1117/12.2208848","DOIUrl":"https://doi.org/10.1117/12.2208848","url":null,"abstract":"Both Optical Coherence Tomography (OCT) and Single Fiber Reflectance Spectroscopy (SFR) are used to determine various optical properties of tissue. We developed a method combining these two techniques to measure the scattering anisotropy (g1) and γ (=1-g2/1-g1), related to the 1st and 2nd order moments of the phase function. The phase function is intimately associated with the cellular organization and ultrastructure of tissue, physical parameters that may change during disease onset and progression. Quantification of these parameters may therefore allow for improved non-invasive, in vivo discrimination between healthy and diseased tissue. With SFR the reduced scattering coefficient and γ can be extracted from the reflectance spectrum (Kanick et al., Biomedical Optics Express 2(6), 2011). With OCT the scattering coefficient can be extracted from the signal as a function of depth (Faber et al., Optics Express 12(19), 2004). Consequently, by combining SFR and OCT measurements at the same wavelengths, the scattering anisotropy (g) can be resolved using µs’= µs*(1-g). We performed measurements on a suspension of silica spheres as a proof of principle. The SFR model for the reflectance as a function of the reduced scattering coefficient and γ is based on semi-empirical modelling. These models feature Monte-Carlo (MC) based model constants. The validity of these constants - and thus the accuracy of the estimated parameters - depends on the phase function employed in the MC simulations. Since the phase function is not known when measuring in tissue, we will investigate the influence of assuming an incorrect phase function on the accuracy of the derived parameters.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"106 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114322231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
To construct a Stochastic Optical Reconstruction Microscopy (STORM) image one should collect sufficient number of localized fluorophores to satisfy Nyquist criterion. This requirement limits time resolution of the method. In this work we propose a probabalistic approach to construct STORM images from a subset of localized fluorophores 3-4 times sparser than required from Nyquist criterion. Using a set of STORM images constructed from number of localizations sufficient for Nyquist criterion we derive a model which allows us to predict the probability for every location to be occupied by a fluorophore at the end of hypothetical acquisition, having as an input parameters distribution of already localized fluorophores in the proximity of this location. We show that probability map obtained from number of fluorophores 3-4 times less than required by Nyquist criterion may be used as superresolution image itself. Thus we are able to construct STORM image from a subset of localized fluorophores 3-4 times sparser than required from Nyquist criterion, proportionaly decreasing STORM data acquisition time. This method may be used complementary with other approaches desined for increasing STORM time resolution.
{"title":"Restoration of STORM images from sparse subset of localizations (Conference Presentation)","authors":"A. Moiseev, G. Gelikonov, V. Gelikonov","doi":"10.1117/12.2212825","DOIUrl":"https://doi.org/10.1117/12.2212825","url":null,"abstract":"To construct a Stochastic Optical Reconstruction Microscopy (STORM) image one should collect sufficient number of localized fluorophores to satisfy Nyquist criterion. This requirement limits time resolution of the method. In this work we propose a probabalistic approach to construct STORM images from a subset of localized fluorophores 3-4 times sparser than required from Nyquist criterion. Using a set of STORM images constructed from number of localizations sufficient for Nyquist criterion we derive a model which allows us to predict the probability for every location to be occupied by a fluorophore at the end of hypothetical acquisition, having as an input parameters distribution of already localized fluorophores in the proximity of this location. We show that probability map obtained from number of fluorophores 3-4 times less than required by Nyquist criterion may be used as superresolution image itself. Thus we are able to construct STORM image from a subset of localized fluorophores 3-4 times sparser than required from Nyquist criterion, proportionaly decreasing STORM data acquisition time. This method may be used complementary with other approaches desined for increasing STORM time resolution.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115132347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
É. De Montigny, N. Goulamhoussen, W. Madore, M. Strupler, A. Maniakas, T. Ayad, C. Boudoux
While thyroidectomy is considered a safe surgery, dedicated tools facilitating tissue identification during surgery could improve its outcome. The most common complication following surgery is hypocalcaemia, which results from iatrogenic removal or damage to parathyroid glands. This research project aims at developing and validating an instrument based on optical microscopy modalities to identify tissues in real time during surgery. Our approach is based on a combination of reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) to obtain multi-scale morphological contrast images. The orthogonal field of views provide information to navigate through the sample. To allow simultaneous, synchronized video-rate imaging in both modalities, we designed and built a dual-band wavelength-swept laser which scans a 30 nm band centered at 780 nm and a 90 nm band centered at 1310 nm. We built an imaging setup integrating a custom-made objective lens and a double-clad fibre coupler optimized for confocal microscopy. It features high resolutions in RCM (2µm lateral and 20 µm axial) in a 500 µm x 500 µm field-of-view and a larger field-of-view of 2 mm (lateral) x 5 mm (axial) with 20 µm lateral and axial resolutions in OCT. Imaging of ex vivo animal samples is demonstrated on a bench-top system. Tissues that are visually difficult to distinguish from each other intra-operatively such as parathyroid gland, lymph nodes and adipose tissue are imaged to show the potential of this approach in differentiating neck tissues. We will also provide an update on our ongoing clinical pilot study on patients undergoing thyroidectomy.
虽然甲状腺切除术被认为是一种安全的手术，但在手术过程中方便组织识别的专用工具可以改善其结果。手术后最常见的并发症是低钙血症，这是由医源性切除或甲状旁腺损伤引起的。本研究项目旨在开发和验证一种基于光学显微镜模式的仪器，用于在手术过程中实时识别组织。我们的方法是基于反射共聚焦显微镜(RCM)和光学相干断层扫描(OCT)的组合来获得多尺度形态对比图像。视图的正交场提供了在样本中导航的信息。为了实现两种模式下的同步视频速率成像，我们设计并制造了一个双频波长扫描激光器，扫描以780纳米为中心的30纳米波段和以1310纳米为中心的90纳米波段。我们建立了一个成像装置，集成了一个定制的物镜和一个优化的双包层光纤耦合器，用于共聚焦显微镜。它具有高分辨率的RCM (2 μ m横向和20 μ m轴向)，500 μ m x 500 μ m视场和更大的视场2mm(横向)x 5mm(轴向)，20 μ m横向和轴向分辨率，在oct中展示了离体动物样品的成像。术中视觉上难以区分的组织，如甲状旁腺、淋巴结和脂肪组织，通过成像显示该方法在区分颈部组织方面的潜力。我们还将提供正在进行的甲状腺切除术患者临床试点研究的最新情况。
{"title":"Simultaneous multi-scale microscopy as a potential dedicated tool for intra-operative parathyroid identification during thyroid surgery (Conference Presentation)","authors":"É. De Montigny, N. Goulamhoussen, W. Madore, M. Strupler, A. Maniakas, T. Ayad, C. Boudoux","doi":"10.1117/12.2211200","DOIUrl":"https://doi.org/10.1117/12.2211200","url":null,"abstract":"While thyroidectomy is considered a safe surgery, dedicated tools facilitating tissue identification during surgery could improve its outcome. The most common complication following surgery is hypocalcaemia, which results from iatrogenic removal or damage to parathyroid glands. This research project aims at developing and validating an instrument based on optical microscopy modalities to identify tissues in real time during surgery. Our approach is based on a combination of reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) to obtain multi-scale morphological contrast images. The orthogonal field of views provide information to navigate through the sample. To allow simultaneous, synchronized video-rate imaging in both modalities, we designed and built a dual-band wavelength-swept laser which scans a 30 nm band centered at 780 nm and a 90 nm band centered at 1310 nm. We built an imaging setup integrating a custom-made objective lens and a double-clad fibre coupler optimized for confocal microscopy. It features high resolutions in RCM (2µm lateral and 20 µm axial) in a 500 µm x 500 µm field-of-view and a larger field-of-view of 2 mm (lateral) x 5 mm (axial) with 20 µm lateral and axial resolutions in OCT. Imaging of ex vivo animal samples is demonstrated on a bench-top system. Tissues that are visually difficult to distinguish from each other intra-operatively such as parathyroid gland, lymph nodes and adipose tissue are imaged to show the potential of this approach in differentiating neck tissues. We will also provide an update on our ongoing clinical pilot study on patients undergoing thyroidectomy.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114663178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Optical coherence tomography (OCT) based microangiography (OMAG) is a new imaging technique enabling the visualization of blood flow within microcirculatory tissue beds in vivo with high resolution. In this talk, the concept and advantages of OMAG will be discussed and its potential clinical applications in the dermatology will be shown, demonstrating its usefulness in the clinical monitoring and therapeutic treatment of various skin pathologies, e.g. acne, port wine stain and wound healing.
{"title":"Optical coherence tomography based microangiography: A tool good for dermatology applications (Conference Presentation)","authors":"Ruikang K. Wang, U. Baran, W. Choi","doi":"10.1117/12.2217625","DOIUrl":"https://doi.org/10.1117/12.2217625","url":null,"abstract":"Optical coherence tomography (OCT) based microangiography (OMAG) is a new imaging technique enabling the visualization of blood flow within microcirculatory tissue beds in vivo with high resolution. In this talk, the concept and advantages of OMAG will be discussed and its potential clinical applications in the dermatology will be shown, demonstrating its usefulness in the clinical monitoring and therapeutic treatment of various skin pathologies, e.g. acne, port wine stain and wound healing.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"37 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127080051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Optical imaging of objects embedded within scattering media such as biological tissues suffers from the strong background noise due to multiple light scattering. The signal strength from the target objects decays exponentially at the length scale of the scattering mean free path, which is typically on the order of 100 micron for biological tissues. As a consequence, targets located at a depth of just a few scattering mean free paths lose their fine details. In this work, we performed synthetic aperture imaging of targets embedded within a scattering medium and demonstrated that the aperture synthesis process can suppress multiple scattering background better than conventional incoherent imaging. In the reflection geometry, we sent planar waves of various incidence angles and recorded the phase and amplitude maps of the reflected waves using off-axis digital holographic microscopy. A He-Ne laser was used as a light source and target objects were sandwiched between scattering layers made of PDMS mixed with polystyrene beads. We converted each reflected images taken at specific incidence angles into the maps of in-plane momentum difference between reflected and incidence waves. We then synthesized the maps in such a way that the scattered waves with the same momentum differences were added together. In this way, single-scattered waves from the targets were added coherently, which made them outgrow the incoherently added multiple-scattered waves. We achieved 1 micron lateral resolution for a target located deeper than four times the scattering mean free path in which conventional incoherent imaging fails to work.
{"title":"Synthetic aperture imaging of objects embedded within scattering media (Conference Presentation)","authors":"Pilsung Kang, W. Choi","doi":"10.1117/12.2211535","DOIUrl":"https://doi.org/10.1117/12.2211535","url":null,"abstract":"Optical imaging of objects embedded within scattering media such as biological tissues suffers from the strong background noise due to multiple light scattering. The signal strength from the target objects decays exponentially at the length scale of the scattering mean free path, which is typically on the order of 100 micron for biological tissues. As a consequence, targets located at a depth of just a few scattering mean free paths lose their fine details. In this work, we performed synthetic aperture imaging of targets embedded within a scattering medium and demonstrated that the aperture synthesis process can suppress multiple scattering background better than conventional incoherent imaging. In the reflection geometry, we sent planar waves of various incidence angles and recorded the phase and amplitude maps of the reflected waves using off-axis digital holographic microscopy. A He-Ne laser was used as a light source and target objects were sandwiched between scattering layers made of PDMS mixed with polystyrene beads. We converted each reflected images taken at specific incidence angles into the maps of in-plane momentum difference between reflected and incidence waves. We then synthesized the maps in such a way that the scattered waves with the same momentum differences were added together. In this way, single-scattered waves from the targets were added coherently, which made them outgrow the incoherently added multiple-scattered waves. We achieved 1 micron lateral resolution for a target located deeper than four times the scattering mean free path in which conventional incoherent imaging fails to work.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127140796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wide-field fluorescent imaging severely suffers low resolution and poor contrast from out-of-focus background to image biological samples. In order to enhance optical sectioning capability, Confocal approach has been developed to filter out-of-focus background using point-to-point detection through a spatial pinhole. Recently, active structured illumination in wide-field fashion has been developed to reduce the transversal scanning cost, but still requires scanning in axial direction. Here, we present a wide-field multi-focal fluorescence microscopy incorporating spatial-spectral volume holographic gratings (MVHGs) with 3D active structured illumination to obtain optically sectioned images without scanning is presented. In contrast to conventional holographic techniques, which in general can not obtain fluorescence images, our approach does not require the formation of a hologram during imaging and is compatible with fluorescence based methods of imaging. Our approach requires pair-wise multi-depth resolved images, one with 3D active illumination, and the other with standard uniform illumination. Our approach is configured such that 3D illuminated planes occur inside the specimen, and also serve as the structured modulation for multiple axial planes imaged by MVHGs and display laterally onto the camera. The system can also be combined with micro-objective and relay systems for endoscopic operation. We demonstrate the proposed system’s ability to simultaneously obtain wide-field, optically sectioned, and multi-depth resolved images of fluorescently labeled tissue structures.
{"title":"Optically sectioned spatial-spectral coded holographic fluorescence microscopy (Conference Presentation)","authors":"Hsi-Hsun Chen, C. Lin, Wei-Tang Lin, Yuan Luo","doi":"10.1117/12.2213937","DOIUrl":"https://doi.org/10.1117/12.2213937","url":null,"abstract":"Wide-field fluorescent imaging severely suffers low resolution and poor contrast from out-of-focus background to image biological samples. In order to enhance optical sectioning capability, Confocal approach has been developed to filter out-of-focus background using point-to-point detection through a spatial pinhole. Recently, active structured illumination in wide-field fashion has been developed to reduce the transversal scanning cost, but still requires scanning in axial direction. Here, we present a wide-field multi-focal fluorescence microscopy incorporating spatial-spectral volume holographic gratings (MVHGs) with 3D active structured illumination to obtain optically sectioned images without scanning is presented. In contrast to conventional holographic techniques, which in general can not obtain fluorescence images, our approach does not require the formation of a hologram during imaging and is compatible with fluorescence based methods of imaging. Our approach requires pair-wise multi-depth resolved images, one with 3D active illumination, and the other with standard uniform illumination. Our approach is configured such that 3D illuminated planes occur inside the specimen, and also serve as the structured modulation for multiple axial planes imaged by MVHGs and display laterally onto the camera. The system can also be combined with micro-objective and relay systems for endoscopic operation. We demonstrate the proposed system’s ability to simultaneously obtain wide-field, optically sectioned, and multi-depth resolved images of fluorescently labeled tissue structures.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"78 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126099429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Morphological assessment of spermatozoa is of critical importance for in vitro fertilization (IVF), especially intracytoplasmic sperm injection (ICSI)-based IVF. In ICSI, a single sperm cell is selected and injected into an egg to achieve fertilization. The quality of the sperm cell is found to be highly correlated to IVF success. Sperm morphology, such as shape, head birefringence and motility, among others, are typically evaluated under a microscope. Current observation relies on conventional techniques such as differential interference contrast microscopy and polarized light microscopy. Their qualitative nature, however, limits the ability to provide accurate quantitative analysis. Here, we demonstrate quantitative morphological measurement of sperm cells using two types of spectral interferometric techniques, namely spectral modulation interferometry and spectral multiplexing interferometry. Both are based on spectral-domain low coherence interferometry, which is known for its exquisite phase determination ability. While spectral modulation interferometry encodes sample phase in a single spectrum, spectral multiplexing interferometry does so for sample birefringence. Therefore they are capable of highly sensitive phase and birefringence imaging. These features suit well in the imaging of live sperm cells, which are small, dynamic objects with only low to moderate levels of phase and birefringence contrast. We will introduce the operation of both techniques and demonstrate their application to measuring the phase and birefringence morphology of sperm cells.
{"title":"Spectral interferometry for morphological imaging in in vitro fertilization (IVF) (Conference Presentation)","authors":"Yizheng Zhu, Chengshuai Li","doi":"10.1117/12.2213070","DOIUrl":"https://doi.org/10.1117/12.2213070","url":null,"abstract":"Morphological assessment of spermatozoa is of critical importance for in vitro fertilization (IVF), especially intracytoplasmic sperm injection (ICSI)-based IVF. In ICSI, a single sperm cell is selected and injected into an egg to achieve fertilization. The quality of the sperm cell is found to be highly correlated to IVF success. Sperm morphology, such as shape, head birefringence and motility, among others, are typically evaluated under a microscope. Current observation relies on conventional techniques such as differential interference contrast microscopy and polarized light microscopy. Their qualitative nature, however, limits the ability to provide accurate quantitative analysis. Here, we demonstrate quantitative morphological measurement of sperm cells using two types of spectral interferometric techniques, namely spectral modulation interferometry and spectral multiplexing interferometry. Both are based on spectral-domain low coherence interferometry, which is known for its exquisite phase determination ability. While spectral modulation interferometry encodes sample phase in a single spectrum, spectral multiplexing interferometry does so for sample birefringence. Therefore they are capable of highly sensitive phase and birefringence imaging. These features suit well in the imaging of live sperm cells, which are small, dynamic objects with only low to moderate levels of phase and birefringence contrast. We will introduce the operation of both techniques and demonstrate their application to measuring the phase and birefringence morphology of sperm cells.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125502988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. Deniz, Stephan M. Jonas, J. Griffin, Michael Hooper, M. Choma, M. Khokha
The etiology of craniofacial defects is incompletely understood. The ability to obtain large amounts of gene sequence data from families affected by craniofacial defects is opening up new ways to understand molecular genetic etiological factors. One important link between gene sequence data and clinical relevance is biological research into candidate genes and molecular pathways. We present our recent research using OCT as a nondestructive phenotyping modality of craniofacial morphology in Xenopus embryos, an important animal model for biological research in gene and pathway discovery. We define 2D and 3D scanning protocols for a standardized approach to craniofacial imaging in Xenopus embryos. We define standard views and planar reconstructions for visualizing normal anatomy and landmarks. We compare these views and reconstructions to traditional histopathology using alcian blue staining. In addition to being 3D, nondestructive, and having much faster throughout, OCT can identify craniofacial features that are lost during traditional histopathological preparation. We also identify quantitative morphometric parameters to define normative craniofacial anatomy. We also note that craniofacial and cardiac defects are not infrequently present in the same patient (e.g velocardiofacial syndrome). Given that OCT excels at certain aspects of cardiac imaging in Xenopus embryos, our work highlights the potential of using OCT and Xenopus to study molecular genetic factors that impact both cardiac and craniofacial development.
{"title":"OCT imaging of craniofacial anatomy in xenopus embryos (Conference Presentation)","authors":"E. Deniz, Stephan M. Jonas, J. Griffin, Michael Hooper, M. Choma, M. Khokha","doi":"10.1117/12.2213448","DOIUrl":"https://doi.org/10.1117/12.2213448","url":null,"abstract":"The etiology of craniofacial defects is incompletely understood. The ability to obtain large amounts of gene sequence data from families affected by craniofacial defects is opening up new ways to understand molecular genetic etiological factors. One important link between gene sequence data and clinical relevance is biological research into candidate genes and molecular pathways. We present our recent research using OCT as a nondestructive phenotyping modality of craniofacial morphology in Xenopus embryos, an important animal model for biological research in gene and pathway discovery. We define 2D and 3D scanning protocols for a standardized approach to craniofacial imaging in Xenopus embryos. We define standard views and planar reconstructions for visualizing normal anatomy and landmarks. We compare these views and reconstructions to traditional histopathology using alcian blue staining. In addition to being 3D, nondestructive, and having much faster throughout, OCT can identify craniofacial features that are lost during traditional histopathological preparation. We also identify quantitative morphometric parameters to define normative craniofacial anatomy. We also note that craniofacial and cardiac defects are not infrequently present in the same patient (e.g velocardiofacial syndrome). Given that OCT excels at certain aspects of cardiac imaging in Xenopus embryos, our work highlights the potential of using OCT and Xenopus to study molecular genetic factors that impact both cardiac and craniofacial development.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"56 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122866693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yimei Huang, H. Lui, Jianhua Zhao, D. McLean, H. Zeng
Photothermolysis induced by femtosecond (fs) lasers may be a promising modality in dermatology because of its advantages of high precision due to multiphoton absorption and deeper penetration due to the use of near infrared wavelengths. Although multiphoton absorption nonlinear effects are capable of precision targeting, the femtosecond laser photothermolysis could still have effects beyond the targeted area if a sufficiently high dose of laser light is used. Such unintended effects could be minimized by real time monitoring photothermolysis during the treatment. Targeted photothermolytic treatment of ex vivo mouse skin dermis was performed with tightly focused fs laser beams. Images of reflectance confocal microscopy (RCM), second harmonic generation (SHG), and two-photon fluorescence (TPF) of the mouse skins were obtained with integrated multimodal microscopy before, during, and after the laser treatment. The RCM, SHG, and TPF signal intensities of the treatment areas changed after high power femtosecond laser irradiation. The intensities of the RCM and SHG signals decreased when the tissue was damaged, while the intensity of the TPF signal increased when the photothermolysis was achieved. Moreover, the TPF signal was more susceptible to the degree of the photothermolysis than the RCM and SHG signals. The results suggested that multimodal microscopy is a potentially useful tool to monitor and assess the femtosecond laser treatment of the skin to achieve microscopic photothermolysis with high precision.
{"title":"Monitoring femtosecond laser microscopic photothermolysis with multimodal microscopy (Conference Presentation)","authors":"Yimei Huang, H. Lui, Jianhua Zhao, D. McLean, H. Zeng","doi":"10.1117/12.2214798","DOIUrl":"https://doi.org/10.1117/12.2214798","url":null,"abstract":"Photothermolysis induced by femtosecond (fs) lasers may be a promising modality in dermatology because of its advantages of high precision due to multiphoton absorption and deeper penetration due to the use of near infrared wavelengths. Although multiphoton absorption nonlinear effects are capable of precision targeting, the femtosecond laser photothermolysis could still have effects beyond the targeted area if a sufficiently high dose of laser light is used. Such unintended effects could be minimized by real time monitoring photothermolysis during the treatment. Targeted photothermolytic treatment of ex vivo mouse skin dermis was performed with tightly focused fs laser beams. Images of reflectance confocal microscopy (RCM), second harmonic generation (SHG), and two-photon fluorescence (TPF) of the mouse skins were obtained with integrated multimodal microscopy before, during, and after the laser treatment. The RCM, SHG, and TPF signal intensities of the treatment areas changed after high power femtosecond laser irradiation. The intensities of the RCM and SHG signals decreased when the tissue was damaged, while the intensity of the TPF signal increased when the photothermolysis was achieved. Moreover, the TPF signal was more susceptible to the degree of the photothermolysis than the RCM and SHG signals. The results suggested that multimodal microscopy is a potentially useful tool to monitor and assess the femtosecond laser treatment of the skin to achieve microscopic photothermolysis with high precision.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114564770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Photoacoustic tomography (PAT) exploits optical contrast and ultrasonic detection principles to form images of absorbed optical energy density within tissue. Based on the photoacoustic effect, PAT directly and quantitatively measures specific optical absorption. A full-ring ultrasonic transducer array based photoacoustic computed tomography (PACT) system was recently developed for small animal whole-body imaging with a full-view detection angle and high in-plane resolution (100 µm). However, due to the band-pass frequency response of the piezoelectric transducer elements, the reconstructed images present bipolar (both positive and negative) pixel values, which is artificial and counterintuitive for physicians and biologists seeking to interpret the image. Moreover, bipolar pixel values hinder quantification of physiological parameters, such as oxygen saturation and blood flow speed. Unipolar images can be obtained by deconvolving the raw channel data with the transducer’s electrical impulse response and applying non-negativity during iteration, but this process requires complex transducer modeling and time-consuming computation. Here, we present a multi-view Hilbert transformation method to recover the unipolar initial pressure for full-ring PACT. Multi-view Hilbert transformation along the acoustic wave propagation direction minimizes reconstruction artifacts during envelope extraction and maintains the signal-to-noise ratio of the reconstructed images. The in-plane isotropic spatial resolution of this method was quantified to 168 μm within a 20 × 20 mm2 field of view. The effectiveness of the proposed algorithm was first validated by numerical simulations and then demonstrated with ex-vivo mouse brain structural imaging and in-vivo mouse wholebody imaging.
{"title":"Multi-view Hilbert transformation in full-ring-transducer-array based photoacoustic computed tomography (Conference Presentation)","authors":"Lei Li, Guo Li, Liren Zhu, J. Xia, Lihong V. Wang","doi":"10.1117/12.2212662","DOIUrl":"https://doi.org/10.1117/12.2212662","url":null,"abstract":"Photoacoustic tomography (PAT) exploits optical contrast and ultrasonic detection principles to form images of absorbed optical energy density within tissue. Based on the photoacoustic effect, PAT directly and quantitatively measures specific optical absorption. A full-ring ultrasonic transducer array based photoacoustic computed tomography (PACT) system was recently developed for small animal whole-body imaging with a full-view detection angle and high in-plane resolution (100 µm). However, due to the band-pass frequency response of the piezoelectric transducer elements, the reconstructed images present bipolar (both positive and negative) pixel values, which is artificial and counterintuitive for physicians and biologists seeking to interpret the image. Moreover, bipolar pixel values hinder quantification of physiological parameters, such as oxygen saturation and blood flow speed. Unipolar images can be obtained by deconvolving the raw channel data with the transducer’s electrical impulse response and applying non-negativity during iteration, but this process requires complex transducer modeling and time-consuming computation. Here, we present a multi-view Hilbert transformation method to recover the unipolar initial pressure for full-ring PACT. Multi-view Hilbert transformation along the acoustic wave propagation direction minimizes reconstruction artifacts during envelope extraction and maintains the signal-to-noise ratio of the reconstructed images. The in-plane isotropic spatial resolution of this method was quantified to 168 μm within a 20 × 20 mm2 field of view. The effectiveness of the proposed algorithm was first validated by numerical simulations and then demonstrated with ex-vivo mouse brain structural imaging and in-vivo mouse wholebody imaging.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"05 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129957235","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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