Microscopic Fluorescence Spectral Image

In order to obtain chemical or biochemical composition, concentration. and coordinate (3C) information of sample observed at the same time, the fluorescence spectral imaging technique is proposed and the microscopic fluorescence spectral imager (MFSI) is developed. In MFSI system, a monochromator is directly coupled to the microscope via a fiber optic adapter which fit into the Epi-fluorescence illuminator port, then the Epi-fluorescence filter cube is redesigned for spectral excitation imaging, and a cooled CCD camera is incorporated as its detector. The software system is programmed using VC++6.0, which is composed of basic image processing module (IPM) and spectral imaging analysis module (SIAM). SIAM has plentiful special function for spectral images, like spectral images fusion, spectral image segmentation, pixel spectrum analysis and so on. Using the MFSI, the FluoCells prepared. slide #2 (Molecular Probe Co. USA), which shows bovine pulmonary artery endothelial cells (BPAEC) that have been stained by three fluorescence probe, is tested and the fluorescence spectral image cube is obtained. Experimental results indicate that using MFSI is an effective approach for chemical imaging.