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Simultaneous Imaging of Multiple Fluorescent Probes in Bio-Cells

机译:生物细胞中多个荧光探针的同时成像

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In order to obtain the full spectrum from 400 to 800 nm of each pixcel of a microscopic image, a unique spectro-imaging system was developed using an image slicer. The image slicer is composed of 100 photo fibers which are arranged in a matrix of 10 × 10 at the entrance and 100 × 1 at the exit. A line of this 100 signals is passed through a glism and projected onto a CCD. This system was applied to the fluorescent imaging of bio-cells. One of the demonstrative examples was simultaneous measurements of the Ca2+ concentration and the pH using of respective fluorescent probes. An electric signal was applied to BY-2 protoplasts and the fluorescent spectrum from 500 nm to 800 nm was measured every 5 s. The spectrum of the BY-2 protoplasts changed in response to the electric signal and the Ca2+ concentration, and the pH changes could be monitored. The wavelength resolution was satisfactory, but the space resolution was still rough in comparison with the usual microscopic systems. Notwithstanding these conditions, we could obtain discrete data from more than several tens of sites in a single-cell or a chain of several cells.
机译:为了获得显微图像的每个像素的400至800 nm的全光谱,使用图像切片器开发了独特的光谱成像系统。图像切片器由100条光纤组成,这些光纤在入口处以10×10的矩阵排列,在出口处以100×1的矩阵排列。这100条信号中的一条线穿过一格并投射到CCD上。该系统被应用于生物细胞的荧光成像。说明性示例之一是使用各个荧光探针同时测量Ca 2+浓度和pH。将电信号施加到BY-2原生质体,每5秒钟测量500 nm至800 nm的荧光光谱。 BY-2原生质体的光谱响应电信号和Ca2 +浓度而变化,并且可以监测pH值变化。波长分辨率令人满意,但是与通常的显微系统相比,空间分辨率仍然很粗糙。尽管有这些条件,我们仍可以从单个单元或多个单元的链中数十个以上的站点中获取离散数据。

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