3D Imaging System of Two-Photon Excitation Laser Scanning Microscopy

Laser scanning microscopy is a powerful tool for analyzing the structure and function of biological specimens. Although there exist numerous commercial laser scanning microscopes, the lack of supporting software system poses a great challenge to three dimension (3D) imaging in the study of two-photon excitation laser scanning microscopy (2PLSM). This paper proposes a complete framework for developing such software system and implements this complex software system with the combination of Lab VIEW and OpenGL. Firstly, the volume data is captured by scanning the specimen in 3D with laser. Secondly, the acquired data is processed with image mean filtering and image interpolation algorithm. Finally, the ray-casting algorithm is adopted to fulfill volume visualization. Based on the proposed framework, we develop an interactive user interface with Lab view to facilitate 3D data acquisition and image reconstruction. In addition, 3D display of the image is achieved using OpenGL, which makes it easier to observe the 3D image from different angles. The imaging results show that the proposed software system successfully images specimens in three dimensions with sub-micrometer resolution.