3D激光扫描共聚焦显微镜计量特性分析及测试

Measurement characteristics analysis and test of 3D laser scanning confocal microscope

为了合理评定激光扫描共聚焦显微镜(Laser Scanning Confocal Microscope,LSCM)的测量性能,提出了一种用于检验LSCM计量特性的测试方法。首先,依据LSCM成像原理,对其技术特点,特别是轴向分辨率和横向分辨率等进行了理论分析;然后对其计量特性的参数指标进行了归纳,并提出相应的性能测试方法。具体包括采用线间隔为微纳米级的网格板等作为标准器,测试LSCM的放大倍数和光学横向分辨率;采用纳米级高度台阶样板测试LSCM的光学轴向分辨率和轴向定位特性;采用激光干涉仪测试样品台工作性能等。实验证明:该方法能够满足当前普遍应用的LSCM的性能测试需求。

In order to evaluate the measurement performance of laser scanning confocal microscope （LSCM） accurately, one measurement approach for checking LSCM was presented. Firstly, based on the imaging principle, the technical characteristics of LSCM, especially the vertical resolution and lateral resolution were analyzed in theory; then main parameters of the measurement characteristics were summarized, and the corresponding performance testing menthod were proposed. Such as the amplification and the lateral optical resolution of the LSCM were tested with the nanometer and sub micronmeter line spacing grid standard plates; the axial optical resolution and the axial positioning characteristics of the LSCM were tested with the nanometer height steps; the performances of the sample stage were tested with the laser interferometer. The experimental results show that this method can meet the current performance requirements of LSCM.