权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Study for the dynamics of the intracellular Ca concentration changes with high spatial and temporal resolution.

研究具有高空间和时间分辨率的细胞内 Ca 浓度变化的动态。

基本信息

批准号：
02557007
负责人：
OHMORI Harunori
金额：
$ 6.14万
依托单位：
National Insitute for Physiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

We have made an instrument for exchanging two excitation wavelengths for fluorescence dye at the vide-rate by utilizing a galvano-mirror device, combining with a CCD TV camera and a frame processor. By this device, we could record fura-2 fluorescence changes at two excitation wavelengths at 33 msec time intervals to monitor intracellular Ca ionic concentration changes. These paired fluorescence image data give the profile of intracellular Ca ionic concentration changes at the rate of 66 msec. The device is made from two part. The first part is the galvano-mirror device itself and switches the two excitation wavelengths. The second part is a control logic circuit for synchronizing the galvano-mirror device with the image recording CCD camera, and the frame processing system. Since a single frame of a video-image is made of two interlacing fields, the CCD camera is shutter opened for only 15msec during the two component fields are superimposed. The reduction of the light intensity on the … More camera-head is compensated by using an image intensifier coupled CCD camera. It was simple to synchronize all three component devices (the galvano-mirror device, the CCD camera, and the frame processor), but the attempt for image integration for improving the signal to noise ratio of the fluorescence images was relatively hard to be achieved. We have overcome the problem by adopting an integrated circuit for dividing a clock rate and by synchronizing the start of the frame integration with the start of galvano-mirror device.We have not yet applied this device extensively for the physiology experiments. However, we have already succeeded in recording intracellular Ca ionic concentration changes from cultured neurons from the brain stem of a chick, in both at the ordinary TV rate imaging mode and at a slower rate mode with the frame integration. This chopper device is basically vibration-free, and it is expected to be applied in the electrophysiology experiments to record intracellular ionic concentration changes simultaneously with the electrical activity of the cell. Less

我们利用电流镜装置，结合CCD电视摄像机和图像处理机，研制了一种荧光染料双激发波长视频转换仪。通过该装置，我们可以在两个激发波长下以33毫秒的时间间隔记录Fura-2荧光的变化，以监测细胞内Ca离子浓度的变化。这些成对的荧光图像数据给出了细胞内Ca离子浓度以66毫秒的速率变化的曲线。该装置由两部分组成。第一部分是电流镜装置本身，并切换两个激发波长。第二部分是控制逻辑电路，用于同步电流镜装置与图像记录CCD相机和帧处理系统。由于视频图像的单个帧由两个隔行扫描场组成，因此在两个分量场叠加期间，CCD相机的快门仅打开15毫秒。光强度的降低， ...更多信息采用像增强器耦合CCD摄像机对摄像头进行补偿。同步所有三个组件设备（电流镜设备、CCD相机和帧处理器）是简单的，但是用于改善荧光图像的信噪比的图像积分的尝试相对难以实现。我们采用了一个时钟分频的集成电路，并使帧积分的开始与电流镜装置的开始同步，从而克服了这个问题，但这种装置还没有广泛地应用于生理学实验。然而，我们已经成功地记录细胞内钙离子浓度的变化，从培养的神经元从一只鸡的脑干，在普通的电视率成像模式和在一个较慢的速度模式与帧积分。这种斩波器装置基本上是无振动的，它有望应用于电生理实验中，以记录细胞内离子浓度的变化与细胞的电活动的同时。少