生物光子学5-02.ppt

从显微镜厚样本成像原理知道, 第j 幅切片图像是所有样本平面与相应点扩散函数的卷积之和, 每一个切片图像中都包含有焦平面信息和散焦信息, 人们希望从光学切片中去除或减弱散焦信息, 以去除结构假象, 提高图像的分辨率。 * * * * * FD：视场光阑；AD：孔径光阑 * Diaphragm，光阑，膜片? * * * * * * * * * * * * * Birefringence 双折射 * Columnar柱状的 * * * blowfly，绿头苍蝇；ferro-silicon alloy，硅铁合金；retardation plate?延迟板，减速板；Photomicrograph，显微照片 * * * Excited state lifetime The excited state of a fluorophore is characterized by a very short half-life, usually on the order of a few nanoseconds. During this brief period, the excited molecules generally relax toward the lowest vibrational energy level within the electronic excited state. The energy lost in this relaxation is dissipated as heat. It is from the resulting relaxed singlet excited state (S1) that fluorescence emission originates. Emission When a fluorochrome molecule falls from the excited state to the ground state, light is often emitted at a characteristic wavelength. The energy of the emitted photon (hnEM) is the difference between the energy levels of the two states (Fig 2), and that energy difference determines the wavelength of the emitted light (lEM). lEM = hc/EEM?, where E = the energy difference between the energy levels of the two states during emission (EM) of light; h = Planck’s constant; c = the speed of light A laser-scanning instrument or a CCD-camera can be used to measure the intensity of the fluorescent light and subsequently create a digital image of the sample. Image analysis makes it possible to view, measure, render, and quantify the resulting image. bovine vascular 牛血管内皮细胞 * Nucleus细胞核 Tubulin微管蛋白 Mitochondrial线粒体 Golgi complex高尔基体复合物 * 荧光滤块是一个综合模块，一般包括一套双色镜DM、激发滤光片EF和截止滤光片BF，还有底座、接口等。 * EMCCD(Electron-Multiplying CCD)增益 ICCD增强型 * Binning是一种图像读出模式，相邻的像元中感应的电荷被加在一起，以一个像素的模式读出。Binning分为水平方向Binning和垂直方向Binning，水平方向Binning是将相邻的行的电荷加在一起读出，而垂直方向Binning是将相邻的列的电荷加在一起读出，Binning这一技术的优点是能将几个像素联合起来作为一个像素使用，提高灵敏度，输出速度，降低分辨率，当行和列同时采用Binning时，图像的纵横比并不改变，当采用2:2Binning，图像的解析度将减少75%。 * 点扩散函数 (point spread function, 简称PSF)。是描述光学系统对点源解析能力的函数。因为点源在