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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅴ, Jan 23-25, 2002, San Jose, USA >Functional Imaging of Living Paramecium by means of Confocal and Two-PhotonExcitation Fluorescence Microscopy
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Functional Imaging of Living Paramecium by means of Confocal and Two-PhotonExcitation Fluorescence Microscopy

机译：共聚焦和双光子激发荧光显微镜对活草履虫的功能成像

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Confocal and Two-photon excitation laser scanning microscopy allow gathering three-dimensional and temporal information from biological systems exploiting fluorescence labeling and autofluorescence properties. In this work we study biological events linked to functionality in Paramecium primaurelia. The internalization of material in ciliated one-celled organisms (protozoa) occurs via different mechanisms, even if most of nutrients, particulate or not, is taken up by food vacuoles formed at the bottom of the oral cavity. The endocytosis of small-sized molecules occurs at the parasomal sacs, located next the ciliar basal bodies. Vital fluorescent dyes (BSA-FITC, WGA-FITC, dextran-Texas Red, cholesteryl-Bodipy) and autofluorescence were used to study formation, movement, and fusion of vesicles during endocytosis and phagocytosis of Paramecium primaurelia. By immobilizing living cells pulsed with food vacuole and endosome markers at successive times after chasing in unlabeled medium, the intracellular movement and fusion of food vacuoles and of endosomes were visualized. A temporal analysis of fluorescence images and the false-color technique were used. Starting from time series or 3D data sets composite images were generated by associating with each originally acquired image a different color corresponding to each sampling point in time and along the z-axis. Second Harmonic Generation Imaging attempts are also outlined.
机译：共焦和双光子激发激光扫描显微镜允许从利用荧光标记和自发荧光特性的生物系统收集三维和时间信息。在这项工作中，我们研究与草履虫的功能相关的生物学事件。纤毛单细胞生物（原生动物）中的物质内部化是通过不同的机制发生的，即使大多数营养物质（无论是否存在颗粒）被口腔底部形成的食物液泡吸收。小分子的内吞作用发生在纤毛囊中，位于纤毛基体的旁边。重要的荧光染料（BSA-FITC，WGA-FITC，右旋糖酐-得克萨斯红，胆甾醇-Bodipy）和自发荧光用于研究草履虫草履虫的内吞作用和吞噬作用期间囊泡的形成，运动和融合。通过在追逐未标记的培养基后连续时间固定用食物泡和内体标记物脉冲的活细胞，可以观察到食物泡和内体的细胞内运动和融合。使用荧光图像的时间分析和伪彩色技术。从时间序列或3D数据集开始，通过将与每个原始采集的图像相关联的不同颜色与每个采样时间点和沿z轴相关联来生成合成图像。还概述了第二谐波产生成像的尝试。

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