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In Vivo Two Photon Excited Fluorescence (TPEF) Detection of Intracellular Microspheres for Cell Tracer Studies in the Cornea

Author: HaoMing
Tutor: RenQiuShi
School: Shanghai Jiaotong University
Course: Biochemistry and Molecular Biology
Keywords: Two-photon induced fluorescence Microspheres Rabbit corneal Fibroblasts
CLC: Q436
Type: Master's thesis
Year: 2010
Downloads: 14
Quote: 0
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Abstract


By detecting implanted in vivo rabbit cornea by two-photon induced fluorescent microspheres labeled corneal cells to establish a new nonlinear imaging methods . And clinical application of the two-photon induced the laser seized eyepiece diagnosis corneal wound healing process to make the exploration . Cultured in vitro to the in vivo rabbit by the blue-green phosphor , and a diameter of about 1 micron fluorescent microspheres labeled rabbit corneal fibroblast cells ( RCF ) was microinjected into the corneal stroma . After the injection of six hours and six days , two-photon induced fluorescence signal and cell morphology were new design Heidelberg two-photon laser seized eyepiece collection . RCF labeled fluorescent specificity after laser confocal microscopy confirmed . Vitro cell morphology and loss of fluorescent markers by fluorescence inverted microscope evaluation . Experimental results show that the in vitro observations show that the fluorescent marker via RCF cells and microspheres overnight culture after cell endocytosis . Rabbit cornea by microinjection into the living body after the two-photon induced fluorescence signal , respectively in 6 hours, and 6 days after the injection is detected out. And in vivo and in vitro observations , experiments show that endocytosis of fluorescent microspheres has not been detected in the extracellular . This study confirmed using nonlinear optical microscopy to detect labeled cells via microinjection into living tissue collected by the feasibility of the two-photon induced fluorescence signal . This method also provides a viable in vivo in situ long-term follow the practice of the living cells , to avoid photobleaching caused by the conventional fluorescence microscope and light-induced cytotoxicity drawbacks , while providing a deeper tissue penetration and higher axial resolution.

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CLC: > Biological Sciences > Physiology > Analyzer physiology ( sensory physiology) > Photoreceptors (visual )
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