Дисертації з теми "Multi-Photon microscope"
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Chuah, Joon Huang. "A multi-pixel CMOS photon detector for the scanning electron microscope." Thesis, University of Cambridge, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.608077.
Повний текст джерелаGuillemant, Marie. "Development of a three-photon microscope for awake and behaving non-human primates." Electronic Thesis or Diss., université Paris-Saclay, 2023. http://www.theses.fr/2023UPASL025.
Повний текст джерелаMulti-photon microscopy has become a standard technique to study the structural and functional activity in mice but it faces obstacles to be applied in larger animals. It would be particularly advantageous to be able to apply it to macaque monkeys, as they are the animal model of choice to understand the neural mechanisms of high-level cognitive functions such as selective attention, working memory and consciousness. One of the main limiting factors for imaging in larger animals is the dura mater. This tough and opaque layer of tissue protects the brain but is so thick in larger animals that it obstructs imaging. It is therefore commonly removed but this leads to a highly invasive and unstable preparation. The main aim of the current work is to investigate the possibility to record functional activity from the cortex of the rhesus macaque monkey through the natural dura.A multi-photon microscopy setup has been designed with a two-photon and a three-photon microscopy optical paths to record from awake macaque monkeys. The repetition rate of the laser is 2MHz which allows a maximum imaging depth inside the cortex of 520µm at 960nm and 715µm at 1300nm with an additional 120µm-thick layer of dura mater at the surface. Resonance-galvo scanning is used to allow a maximal frame rate of 15.6Hz at a field of view of 620x630µm². In addition to the setup, surgical implants have been developed for long-term and awake imaging.Using an ex vivo study of dura mater from a macaque monkey, the induced optical aberrations are studied by measuring the decrease in spatial resolution of the setup for a varying thickness of dura mater. This reveals that it has no significant impact on the spatial resolution for a thickness up to 150µm at 1300nm. The effective attenuation length of the dura mater is estimated to be 56.5±10.1µm at 960nm and 80.7±5.3µm at 1300nm. These measurements are used to model the maximum imaging depth that can be reached according to the repetition rate of the laser and the thickness of the dura.This model is adjusted and validated using in vivo data from two non-human primates. The effective attenuation length of the natural dura mater and of a regrowth of tissue following a durectomy (called a 'neomembrane') are investigated. Functional recordings have been performed in mice and preprocessed using Suite2P. Viral injection parameters have been tested in three macaque monkeys and we have so far recorded the in vivo structural and functional activity of neurons in one. Finally, the comparison between the use of two- and three-photon microscopy to study non-human primates is discussed. In conclusion, we have set up and optimized a multi-photon microscope for long-term awake imaging of the cortex of non-human primates and shown that it was possible to record down to over 700µm into the cortex (which corresponds to the layers L2/L3) while imaging through the natural dura mater or a neomembrane
Mansfield, Jessica. "Multi-photon microscopy of cartilage." Thesis, University of Exeter, 2008. http://hdl.handle.net/10036/42345.
Повний текст джерелаTsikritsis, Dimitrios. "Vibrational spectroscopy and microscopy in colorectal cancer." Thesis, University of Edinburgh, 2018. http://hdl.handle.net/1842/33049.
Повний текст джерелаRomijn, Elisabeth Inge. "Development of 3-D Quantitative Analysis of Multi-Photon Microscopy Images." Thesis, Norges teknisk-naturvitenskapelige universitet, Institutt for fysikk, 2012. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-18425.
Повний текст джерелаChalal, Mohand. "Structure multi-échelle et propriétés physico-chimiques des gels de polymères thermosensibles." Phd thesis, Université de Grenoble, 2011. http://tel.archives-ouvertes.fr/tel-00680076.
Повний текст джерелаGasecka, Alicja. "Microscopie non-lineaire polarimetrique dans les milieux moleculaires et biologiques." Phd thesis, Université Paul Cézanne - Aix-Marseille III, 2010. http://tel.archives-ouvertes.fr/tel-00560415.
Повний текст джерелаGuiet, Romain. "Étude des mécanismes cellulaires et moléculaires de la migration des macrophages humains dans des environnements en trois dimensions." Toulouse 3, 2011. http://thesesups.ups-tlse.fr/1487/.
Повний текст джерелаTissue infiltration of macrophages is an aggravating factor in many diseases such as chronic inflammation and cancer. Macrophages that infiltrate tumors are called tumor-associated macrophages (TAMs). They promote tumor growth, angiogenesis, invasion and metastasis. Thus, inhibition of macrophage infiltration has become a therapeutic goal. Recently, the team demonstrated that macrophages use the amoeboid (depending on ROCK) or the mesenchymal (depending on proteases) migratory mode according to the extracellular matrix (ECM) architecture in three dimensions (3D). In addition, the study of the mesenchymal migration mode showed that it is dependent on Hck (a phagocyte-specific tyrosine kinase) and its ability to reorganize podosomes (ECM-degrading actin-rich structures) into rosettes. My thesis project was organized around two axes 1) the identification of substrates of Hck and the characterization of their role in the organization of podosomes and 3D migration of macrophages, and 2) the study of the 3D migration mechanisms of primary human monocytes/ macrophages within an in vitro tumor model: tumor cell spheroids. By a proteomic approach, I have identified potential partners and substrates of Hck, including the protein Filamin A (FLNa), a protein interacting with the actin cytoskeleton and integrins. Using different tools (recombinant proteins, antibodies, shRNA. . . ) I showed that: 1) Hck phosphorylates FLNa in vitro, 2) FLNa is localized to podosomes and is necessary for their organization as rosettes under the control of Hck, 3) the podosomes of FLNa-deficient cells have a shorter life span, and 4) the expression of FLNa is required for mesenchymal migration, but not for amoeboid migration of macrophages in a 3D ECM. Thus, FLNa could be a substrate of Hck necessary for the formation and stabilization of podosomes and their organization as rosettes, and is required for the mesenchymal migration of macrophages. In parallel, I developed a model of tumor cell spheroids, which allowed me to show that the infiltration of monocytes or macrophages in this in vitro tissue model of tumor is dependent on ROCK and proteases, signature of the use of the two migration modes. Then, when spheroids were embedded into ECM, I demonstrated that the presence of macrophages infiltrated into the spheroids is necessary to trigger the invasiveness of tumor cells. Indeed, macrophages infiltrate first the surrounding ECM and tumor cells follow macrophages in the matrix outside of the spheroid. Hck-/- macrophages, that are defective in mesenchymal migration, are significantly less effective in promoting the invasion of tumor cells. These results indicate that the activity of migration and matrix remodeling exerted by macrophages is prominent in tumor invasion. These results have established the migratory mode of macrophages infiltrating an in vitro tumor model and a mechanism required for tumor invasiveness promoted by macrophages. Thus during my thesis, I characterized the molecular and cellular mechanisms of 3D migration of human macrophages. Indeed, I have been able to: 1) identify a protein necessary for the mesenchymal migration of macrophages, 2) highlight the use by macrophages of the amoeboid and mesenchymal migration modes during their infiltration into an in vitro tumor model in 3D and 3) show that the matrix remodeling activity of macrophages during their migration plays a critical role in tumor cell invasion
Leclerc, Pierre. "Développement d’un endomicroscope multiphotonique à deux couleurs pour l’imagerie du métabolisme énergétique cellulaire." Thesis, Limoges, 2017. http://www.theses.fr/2017LIMO0053/document.
Повний текст джерелаNonlinear microscopy is a cutting edge imaging modality leading to remarkable step forward in biology but also in the clinical field. To use it at its full potential and at the very heart of clinical practice, there has been several development of fiber-based micro-endoscope. The application for those probes is now limited by few major restrictions, such as the impossibility to collect auto-fluorescence signal from tissues theses being inherently weak such as the fluorescence from NADH or FAD. This limitation reduces the usefulness of the micro-endoscope effectively restraining it to morphological imaging modality requiring staining of the tissue. Our aim is to go beyond this limitation, showing cellular metabolism monitoring, in real time, without any staining. The experimental setup is an upgrade of our precedent one where the reflection- based Grism stretcher is replace with a new generation transmission-based Grism stretcher. Another Laser was also added in order to tune the first laser at 860nm to allow FAD imaging and the second one to 760nm for NADH. The results prove that we assess and image the level of NADH and FAD at subcellular resolution through a five-meter-long fiber. Thus we demonstrate that we are capable of measuring the optical redox ratio in a micro-endoscopic configuration
Liu, Li. "Roles of PMCA Isoforms in Ca2+-Homeostasis and Contractility of Bladder Smooth Muscle: Evidence from PMCA Gene-Ablated Mice." Cincinnati, Ohio : University of Cincinnati, 2007. http://rave.ohiolink.edu/etdc/view.cgi?acc_num=ucin1178307168.
Повний текст джерелаAdvisor: Richard J. Paul. Title from electronic thesis title page (viewed Apr. 4, 2009). Keywords: PMCA (human gene symbols; ATP2B); SERCA2 (human gene symbols; ATP2A2); NCX; bladder smooth muscle; Ca²⁺ homeostasis; gene-altered mice. Ca²⁺ waves; Ca²⁺ sparks; Fura-PE3; Fluo-4; Indo-1; multi-photon microscopy. Includes abstract. Includes bibliographical references.
Ye, Rai-Teng, and 葉睿騰. "Visualizing the spatiotemporal dynamics of corneal endothelial healing by multi-photon microscope." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/kg5wap.
Повний текст джерела國立臺灣大學
醫學工程學研究所
107
The corneal endothelium is a monolayer of hexagonal cells derived from the neural crest and is responsible for maintaining the dehydration of the cornea. Due to the limited proliferative capacity, impaired mature endothelial cells ultimately results in corneal decompensation and loss of vision. However, the cell dynamics of corneal endothelium during homeostasis and regeneration remain unclear. In this study, we characterize that the N-cadherin is the specific marker of corneal endothelial cells. Using multi-photon live imaging, the cell density and nucleus volume in corneal endothelium significantly decreases from the center to periphery. In addition, the corneal endothelial cells are quiescent, arrested at G1 phase, without migration for consecutive 3 days. We use laser ablation with precise spatiotemporal control to create a 50x50μm2 wound in corneal endothelium in vivo. Here we show that the wound healing process can be divided into three phases: latent phase, migration phase, and remodeling phase. The latent phase initiates at 6 hours and followed by the migration phase from 24 hours. The wound approaches closed at 38 hours. Interestingly, our data suggest that corneal endothelial cells detach from the Descemet’s membrane in both the latent phase and remodeling phase. This study not only develops a novel corneal endothelial injury model but also elucidates the wound healing process in vivo. Additionally, the cell loss from Descemet’s membrane during regeneration enables us to validate the new insight into ophthalmic therapy.
Reddy, Gaddum Duemani. "A multi-photon microscope for three dimensional functional recording of fast neuronal activity." Thesis, 2007. http://hdl.handle.net/1911/20637.
Повний текст джерелаPaudel, Hari Prasad. "Coherent beam control through inhomogeneous media in multi-photon microscopy." Thesis, 2015. https://hdl.handle.net/2144/13674.
Повний текст джерелаYang, Po-Lun, and 楊博綸. "Early diagnosis of acute mesenteric ischemia based on multi-photon fluorescence microscopy." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/8d9etz.
Повний текст джерела國立臺灣大學
醫學工程學研究所
106
We have observed the yellow fluorescence of mesenteric ischemia, and ischemia-reperfusion rats with multi-photon fluorescence lifetime and intensity, which based on a femtosecond Ti:sapphire laser. With lifetime trace, single photon, two-photon fluorescence spectrum, and two-photon fluorescence excitation spectrum, we found out that intensity of yellow fluorescence increases during ischemia state, and recovered gradually after reperfusion. The characteristic change of the autofluorescence intensity ratio in two-photon excitation spectrum, which observed only in the ischemia state instead of normal state or reperfusion state. We compared fluorescence decay traces of preishemia, ischemia state, and ischemia-reperfusion state, the results showed these traces are almost overlapping, which indicated that there is no any new fluorescent substance producing during the whole process. From above results, the elevating intensity of ischemia state is due to the concentration of original fluorescent substances increase. We tried to identify that fluorescent substance by High-performance Liquid Chromatography-Mass Spectrophotometer (HPLC-MS), and the preliminary result is riboflavin. With our study result, we believe two-photon fluorescence excitation spectrum, a non-invasive way, is potential to serve as a tool for early diagnosis of acute mesenteric ischemia.
Chen, Ya-Ting, and 陳亞婷. "The research of multi-fluorescence bio-specimens with two-photon fluorescence hyperspectral microscopy." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/kzv338.
Повний текст джерела國立中央大學
光電科學與工程學系
103
The development of biomedical engineering has gradually improved, and molecular imaging has been one of its most important techniques. To solve the limitation of a conventional microscope that merely detects the morphology of samples, we’ve combined hyperspectral imaging with two photon fluorescence microscopy and designed the setup of two photon fluorescence hyperspectral microscopy. This neoteric technique includes the advantages of both microscopies, providing deeper depth penetration, less photo damage to sample, higher axial resolution, and the addition of receiving spectral information of the targeted sample simultaneously. With these properties, it’s more suitable for biomedical research in vivo, it also provides more information of the samples to researchers, and reduces the misjudgment probabilities of researches that are dependent on the morphology. Using the previously developed system, a non-de-scanned two-photon fluorescence hyperspectral microscope with parallel recording, experiments of cells and sectioning skin samples were carried out to test the efficiency of the system. Furthermore, the major aim is to try to actually apply this microscopy to biological researches and to overcome the limitation of multiple fluorescence labeling samples. Multispectral images might generate crosstalk and increase the difficulty of fluorescence discrimination. Therefore, we applied the applicably spectral analysis, linear unmixing, to improve the spectral discriminating ability. In the biological research, there were two parts of experiments. In the first part, cultured cell lines were labeled with multiple fluorescence dyes and the hyperspectral images of these cells were analyzed to show the ability and correctness of linear unmixing. In the second part, mouse skin tissues with hair follicles being fluorescently labeled were used as samples. According to the experimental results and the spectral analyses, the fluorescence and second harmonic generation signals was easily separated. The unexpected signal sources, second harmonic generation, can be removed to avoid influencing doctor’s diagnoses. The two-photon hyperspectral imaging combined with linear unmixing has shown its ability to biomedical fields.
Hsieh, Chien-Tai, and 謝建泰. "Diagnosis of HCC by using a multi-photon fluorescence microscopy of spectrum, intensity, and lifetime." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/47614149878320542315.
Повний текст джерела國立臺灣大學
醫學工程學研究所
99
With multi-photon fluorescence spectra, multi-photon nonlinear microscopy and FLIM images, we successfully build up a bilirubin molecular imaging platform and imaged the metabolic disorders of bilirubins in HCC. By using an infrared femtosecond Cr:forsterite laser source with its wavelength away from two-photon resonance of most endogenous pigments, the distribution and amount of bilirubins can be semi-quantitively mapped over low-background autofluorescences. For non-tumor regions of liver tissues, the autofluorescences of bilirubins are strong and emitted from granular vesicles in hepatocytes. In poorly-differentiated HCC, the granular vesicles which emit fluorescence disappear and the intensity of multi-photon fluorescence is apparently reduced. If the hepatocytes have cholestasis, which is the signature of well-differentiated HCC, the fluorescence intensity were much higher than non-tumor part and the spectra showed an extra 800nm peak. These results indicate that, based on the multiphoton microscopy of autofluorescence spectra, intensities, and lifetimes, endogenous bilirubins are useful markers to indicate the presence and the staging of HCC. Without an extra labeling or chemical assays, disorder of bilirubins metabolisms in HCC can be sensitively detected.
Lo, Shu-Yen, and 羅書硯. "Utilizing intravital multi-photon microscopy to monitor the effects of fatty liver disease on hepatobiliary metabolism." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/4u5n9c.
Повний текст джерела國立臺灣大學
物理研究所
98
Liver disease represent a major health problem worldwide. Fatty liver is a common liver disease which can progress to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis. In this thesis, in order to elucidate the effects of fatty liver on hepatobiliary metabolism, we combined multi-photon microscopy with a home-built intravital hepatic imaging chamber (IHIC) to image and quantify the metabolic capacity of liver in methionine- and choline-deficient (MCD) dietary animal model. Mice were imaged at 2, 4, 6, and 8 weeks after they were first fed with MCD. Non-fluorescent 6-carboxyfluorescein diacetate (6-CFDA) which is hydrolyzed by the hepatocytes into fluorescent 6-carboxyfluorescein (6-CF) and excreted into the bile canaculi by the multidrug resistant proteins (mrps) is then used as the molecular probe in tracing the effects of fatty liver on hepatobiliary metabolism. Our data showed that the metabolic function of liver in fatty liver diseased mice is impaired in comparison with that of normal mice.
Chang, Po-Shou. "Using Intravital Multi-photon Fluorescence Microscopy in the Investigation of Mouse Liver Inflammation Induced by Lipopolysaccharide(LPS)." 2008. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-2201200817181600.
Повний текст джерелаChang, Po-Shou, and 張栢壽. "Using Intravital Multi-photon Fluorescence Microscopy in the Investigation of Mouse Liver Inflammation Induced by Lipopolysaccharide(LPS)." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/18875343357039961701.
Повний текст джерела臺灣大學
物理研究所
96
The parenchyma of liver has a highly organized and densely crowded sinusoidal system. They are tiny capillary-like vessels, therefore leukocytes and other blood cells must traverse through these vessels in contact with the endothelial cells flanking the sinusoids. By the use of intravital two-photon microscopy and hepatic imaging chamber we can study the response in vivo after administration of lipopolysaccharide (LPS), which is commonly used to model inflammatory mechanism. Whereas it is a critical and apparent process of the inflammatory response that the leukocytes recruitment at the inflamed tissues, these leukocytes roll along the vascular wall until they encounter stimulus produced by the endothelium cells and macrophages. The fluorescence micrographs of the hepatic microcirculation show that leukocytes accumulate within the sinusoids, then transmigrate across endothelial layer and into the inflammatory hepatocytes. We can also observe the metabolism of 6-CFDA within hepatocytes and the subsequent excretion in bile canaliculi.
Tang, Zong-Han, and 湯宗翰. "Utilizing multi-photon microscopy to study mechanism of septic cell shedding in small intestine of mice in vivo." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/26022588534024301082.
Повний текст джерела國立臺灣大學
物理研究所
99
Epithelium of the small intestine, a mono discontinuous layer which consists of many epithelial cells but few gaps, plays a critical role in supporting gut barrier function. These epithelial cells function by migrating and shedding, that is, cells are shed from villi and create gaps. Cell shedding and migration are closely related to the defect of barrier function. However, the connection between abnormality of cell shedding and serious disease such as sepsis is not clear. This study used multi-photon microscopy to understand the mechanism of septic cell shedding in vivo. We constructed a sepsis model by lipopolysaccharide (LPS) and a dangerous sepsis model by LPS puls burn. Finally, the motion of cell shedding and migration was directly observed and detail was completely presented. We found in normal model cells formed dynamic equilibrium of shedding and production. However, in dangerous sepsis model epithelium presented intense shedding and increased permeability.
Purnapatra, Subhajit Banergjee. "Spatial Filtering Techniques for Large Penetration Depth and Volume Imaging in Fluorescence Microscopy." Thesis, 2013. http://etd.iisc.ernet.in/2005/3351.
Повний текст джерелаSchrandt, Christian John. "Chronic monitoring of cortical hemodynamics after ischemic stroke using funcional optical imaging techniques." Thesis, 2015. http://hdl.handle.net/2152/30334.
Повний текст джерелаtext
Freeman, Kim Renee. "In situ three-dimensional reconstruction of mouse heart sympathetic innervation by two-photon excitation fluorescence imaging." Thesis, 2014. http://hdl.handle.net/1805/4030.
Повний текст джерелаThe sympathetic nervous system strongly modulates the contractile and electrical function of the heart. The anatomical underpinnings that enable a spatially and temporally coordinated dissemination of sympathetic signals within the cardiac tissue are only incompletely characterized. In this work we took the first step of unraveling the in situ 3D microarchitecture of the cardiac sympathetic nervous system. Using a combination of two-photon excitation fluorescence microscopy and computer-assisted image analyses, we reconstructed the sympathetic network in a portion of the left ventricular epicardium from adult transgenic mice expressing a fluorescent reporter protein in all peripheral sympathetic neurons. The reconstruction revealed several organizational principles of the local sympathetic tree that synergize to enable a coordinated and efficient signal transfer to the target tissue. First, synaptic boutons are aligned with high density along much of axon-cell contacts. Second, axon segments are oriented parallel to the main, i.e., longitudinal, axes of their apposed cardiomyocytes, optimizing the frequency of transmitter release sites per axon/per cardiomyocyte. Third, the local network was partitioned into branched and/or looped sub-trees which extended both radially and tangentially through the image volume. Fourth, sub-trees arrange to not much overlap, giving rise to multiple annexed innervation domains of variable complexity and configuration. The sympathetic network in the epicardial border zone of a chronic myocardial infarction was observed to undergo substantive remodeling, which included almost complete loss of fibers at depths >10 µm from the surface, spatially heterogeneous gain of axons, irregularly shaped synaptic boutons, and formation of axonal plexuses composed of nested loops of variable length. In conclusion, we provide, to the best of our knowledge, the first in situ 3D reconstruction of the local cardiac sympathetic network in normal and injured mammalian myocardium. Mapping the sympathetic network connectivity will aid in elucidating its role in sympathetic signal transmisson and processing.