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    Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/64414

    Title: Calibrating z distance for confocal microscope
    Authors: 王筑萱;Wang,Chu-hsuan
    Contributors: 生物物理研究所
    Keywords: 共軛焦顯微鏡;z軸校正;折射率不匹配;confocal microscope;z calibration;index mismatch
    Date: 2014-06-03
    Issue Date: 2014-08-11 18:15:07 (UTC+8)
    Publisher: 國立中央大學
    Abstract: 共軛焦顯微鏡已被廣泛運用於建立三維模型中,它藉由在不同垂直方向的位置拍攝一系列堆疊的二維影像來建立模型,但是尚未有基於影像特徵來精確測量三維的距離的方法,要藉由影像來確認垂直方向的移動距離是更為困難。而且當鏡頭的介質與樣品的浸泡液體折射率不相同時,在拍攝影像時會遇到許多問題,像是焦平面移動、像差、吸收與散射,當我們拍攝較深的影像時這些現象會造成影像的變形和亮度的降低。

    ;Confocal microscopes have been widely used to produce a 3D image by acquiring stacks of 2D images. Yet to have accurate 3D distance measurement between features based on images, it is necessary to calibrate pixel-to-micron conversation, which is especially difficult along the optical axis (z direction). When there is a refractive index difference between immersion medium of a sample and an objective, there are problems such as focus shift, aberrations, absorption and scattering which contribute to dramatic reduction in signal intensity as the focus gets deeper into the sample. In this thesis, I will look into the effects of refractive index on 3D imaging with the emphasis on z distance calibration. We employ a simple device which consists of a 2D periodic and fluorescent pattern mounted at 60 degrees with respect to z direction. We can easily compare the interval distance between the original and z-projected image to compute x, y, and z conversion factors and also intensity patterns. We systematic change the immersion medium index and find the relationship between z-conversion factor, intensity drop, and index-mismatch.
    Appears in Collections:[生物物理研究所 ] 博碩士論文

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