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Cell-dynamics Analyzing Apparatus with Multispectral Fluorescence Images Obtained by a Cooled-CCD Camera.

具有通过冷却 CCD 相机获得的多光谱荧光图像的细胞动力学分析装置。

基本信息

批准号：
01870007
负责人：
TAKAHASHI Kunitaro
金额：
$ 6.98万
依托单位：
Faculty of Medicine, University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B).
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

This project aimed to design and make the multispectral image acquiring system consisting of an epifluorescence microscope, a pulse-motor driven interference-filter exchanger, a cooled-CCD camera, and a 32 bit personal computer. The filter exchanger apparatus was designed to insert one out of four interference filters alternatively into the relatively long parallel optical path designed to be placed between the object and eyepiece. The four filters were selected from a set of 11 bandpass filters with half transmittance width 20 nanometer covering the wave length range from 400 to 600 nanometer. The exchange was driven by a computer-controlled pulse motor for which commands were delivered through the interface of the personal computer. The fluorescence images were acquired through the noise-less, ultra-widely dynamically ranged and ultra-highly sensitive cooled-CCD camera. The image acquisition was performed by the commands from the personal computer through the GPIB interface. The embryonic preparation injected with multiple fluorescent dyes or fixed and stained with fluorescent probes were used for obtaining multiple spectral images with this system. The obtained images were analyzed by the programs to extract the material images from the multispectral images, such as proposed by Kawata et al. The programs performed the calculation of fluorescence intensity matrix. The eigen values of the matrix represented the ratios of principal components and the values helped to determine the number of the component chemicals which were responsible for the multispectral images. Then, knowing the spectra of the chemicals, it was possible to reconstruct the material images from multispectral images.

该项目旨在设计和制作多光谱图像采集系统，该系统由落射荧光显微镜、脉冲电机驱动的干涉滤光片交换器、冷却 CCD 相机和 32 位个人计算机组成。滤光器交换装置被设计成将四个干涉滤光器中的一个交替插入到被设计为放置在物体和目镜之间的相对较长的平行光路中。这四个滤光片是从一组 11 个带通滤光片中选出的，半透射率宽度为 20 纳米，覆盖了 400 至 600 纳米的波长范围。该交换由计算机控制的脉冲电机驱动，命令通过个人计算机的接口传送。荧光图像是通过无噪声、超宽动态范围和超高灵敏度的冷却 CCD 相机获取的。图像采集是通过个人计算机通过 GPIB 接口发出的命令来执行的。注射多种荧光染料或用荧光探针固定和染色的胚胎制剂用于通过该系统获得多个光谱图像。通过程序对获得的图像进行分析，以从多光谱图像中提取材料图像，例如 Kawata 等人提出的方法。该程序执行荧光强度矩阵的计算。矩阵的特征值代表主成分的比率，这些值有助于确定造成多光谱图像的化学成分的数量。然后，了解化学物质的光谱，就可以从多光谱图像重建材料图像。