Design Of The Software In The Photoelectric Associated Detection System To Measure [Ca～(2+)]i In Living Cells

As an important intracellular secondary messenger, Ca²⁺ regulates many physiological activities of cells. So it is important to research the activity disciplinarian of Ca²⁺ inside and outside cellular. Fluorescence microscopy is a primary method to detect [Ca²⁺]i in living cells for the advantages of high temporal resolution, and less phototoxicity to the cells. Therefore, the design of the fluorescent microscopy system to detect [Ca²⁺]i in living cells is to be an effective tool for the research of physiology and biology. The primary work mentioned in this article is to develop the control and analysis software in the fluorescence microscopy system to detect [Ca²⁺]i in living cells. The main function of the software is to control the monochromator to emit monochromatic light, which is led to fluorescent microscope by fiber and is focused on the specimen plane to excitated the fluorescence of the fluorescent probe within the cell; then the fluorescent signal is detected by the photomultiplier tube and is transferred to electric signal, which is collected by data acquisition and sent to PC; at last the electric signal is stored and computed by the software to get the [Ca²⁺]i. The research process of the software has been discussed and the design target, principle, research method and the procedure of debug and experimentation are introduced. This software is an MFC-based application of Windows operating system. The object-oriented programming method and the multithread technology are used in the program, and ABF (Axon Binary File) format is used to save original data acquired by data acquisition. The use of the above advanced method and technology has improved developing efficiency greatly and featured good reusability and maintainability of the software. At last, we use the software to measure the [Ca²⁺]i in living β-cell, and the results demonstrate that the software can control the monochromator to emit monochromatic light and to keep the time of exposure accurately, and to measure the [Ca²⁺]i in living cells. It has been proved to be an effective tool for the study in cellular biophysics.