Lateral resolution testing of a novel developed confocal microscopic imaging system

Xin Zhang; Yunhai Zhang; Jian Chang; Wei Huang; Xiaojun Xue; Yun Xiao

doi:10.1117/12.2202799

15 October 2015 Lateral resolution testing of a novel developed confocal microscopic imaging system

Xin Zhang, Yunhai Zhang, Jian Chang, Wei Huang, Xiaojun Xue, Yun Xiao

Proceedings Volume 9672, AOPC 2015: Advanced Display Technology; and Micro/Nano Optical Imaging Technologies and Applications; 96720W (2015) https://doi.org/10.1117/12.2202799
Event: Applied Optics and Photonics China (AOPC2015), 2015, Beijing, China

Abstract

Laser scanning confocal microscope has been widely used in biology, medicine and material science owing to its advantages of high resolution and tomographic imaging. Based on a set of confirmatory experiments and system design, a novel confocal microscopic imaging system is developed. The system is composed of a conventional fluorescence microscope and a confocal scanning unit. In the scanning unit a laser beam coupling module provides four different wavelengths（405nm，488nm，561nm and 638nm）which can excite a variety of dyes. The system works in spot-to-spot scanning mode with a two-dimensional galvanometer. A 50 microns pinhole is used to guarantee that stray light is blocked and only the fluorescence signal from the focal point can be received . The three-channel spectral splitter is used to perform fluorescence imaging at three different working wavelengths simultaneously. The rat kidney tissue slice is imaged using the developed confocal microscopic imaging system. Nucleues labeled by DAPI and kidney spherule curved pipe labeled by Alexa Fluor 488 can be imaged clearly and respectively, realizing the distinction between the different components of mouse kidney tissue. The three-dimensional tomographic imaging of mouse kidney tissue is reconstructed by several two-dimensional images obtained in different depths. At last the resolution of the confocal microscopic imaging system is tested quantitatively. The experimental result shows that the system can achieve lateral resolution priority to 230nm.

Citation Download Citation

Xin Zhang, Yunhai Zhang, Jian Chang, Wei Huang, Xiaojun Xue, and Yun Xiao "Lateral resolution testing of a novel developed confocal microscopic imaging system", Proc. SPIE 9672, AOPC 2015: Advanced Display Technology; and Micro/Nano Optical Imaging Technologies and Applications, 96720W (15 October 2015); https://doi.org/10.1117/12.2202799

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KEYWORDS

Confocal microscopy

Imaging systems

Luminescence

Kidney

Microscopes

Image resolution

Tissues

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