The Design And Application Of Fluorescent Microscopy Monochromator System

As the important intracellular secondary messenger, Ca2+ regulates many physiological activities of cells. The [Ca2+]i detecting system, which is based on the fluorescence microscopy, is an important experiment equipment in biophysics research. But the technical research and manufacture of monochromator are lagged in our country. This article gives a design to monochromator by simulating and analyzing the mathematical model, and the demo device has been used in the [Ca2+]i detecting experiment.The monochromator uses short-arc Xenon lamp as the light source. All the light is converged to a point light source by the elliptical mirror. During the course of collimation, grating diffraction, focus and coupling, the monochromatic light output from the optical fiber. The light power and spectral purity are the tow technical essentials of the monochromator. Using mathematical model to simulate the monochromator system, the optical and mechanical design could be made by the emulation result. After calibration, the monochromator is used in [Ca2+]i detecting experiment and gets eligible result.The research content in this paper could be divided into three parts.The first part introduces the principle and structure of monochromator, the choosing and design methods of the optical components.The second part analyses the output power and spectral purity by mathematical model. The simulation result gives theoretic reasons for optical system design. This part also introduces the work principle of diffraction grating.The third part narrates how to calibrate all parts of the [Ca2+]i detecting system. Using the system, the dynamic curve of [Ca2+]i in PC12 cells after high K+ depolarization is given.