论文部分内容阅读
为了研究折射率不匹配引入的像差对共聚焦显微镜成像的影响,根据Richards和Wolf矢量衍射积分建立计算模型,分析共聚焦显微镜常用的两种情况中存在折射率不匹配时的点扩散函数(PSF),并通过实验进行验证。光束经过折射率不匹配的油-盖玻片-水聚焦时,探测深度从0.5μm增至10μm,其PSF的z轴半峰全宽(FWHM)从0.4391μm变为0.931μm,其光强最大值从1.161降为0.4119。当探测深度为10μm时,折射率匹配的空气-空气-空气的PSF的z轴FWHM为1.059μm,光强最大值为1;而折射率不匹配的空气-玻璃-水的PSF的z轴FWHM为7.5μm,光强最大值为0.04632。当探测深度较大时,折射率不匹配引入的像差会导致光斑变大和不对称,能量分散,影响成像质量。
In order to study the influence of aberrations introduced by refractive index mismatch on confocal microscopy imaging, a calculation model based on Richards and Wolf vector diffraction integral was established, and the point spread function (refractive index mismatch) in the two cases commonly used in confocal microscopy PSF), and verified by experiment. When the beam is focused by the refractive index mismatch oil-cover glass-water, the detection depth increases from 0.5μm to 10μm. The FWHM of the PSF changes from 0.4391μm to 0.931μm, The value dropped from 1.161 to 0.4119. The z-axis FWHM of refractive index-matched air-air-air PSF is 1.059 μm with a maximum light intensity of 1 at a probe depth of 10 μm and the z-axis FWHM of PSF with refractive index mismatch 7.5μm, the maximum light intensity of 0.04632. When the detection depth is large, the refractive index mismatch introduced into the aberration will lead to larger and asymmetric spot, the energy dispersion, affecting the imaging quality.