Information processing of multidimensional simultaneity fluorescence with two-photon array excitation

Abstract

Microscopical imaging method of five-dimensional simultaneity fluorescence information is a new technology for acquiring fluorescence information, which makes use of two-photon array excitation to acquire information of fluorescence position, fluorescence spectrum and fluorescence lifetime for each array element. It removes the limitation on existing fluorescence detection technology in which different functional informations are not obtained simultaneously. In this paper, the methods for deriving geometrical intensity image, different spectral intensity images and different spectral life-time images from the complex information of the new technology are presented. Because of array excitation, there is asymmetry in the array excitation intensity, which influences the excited fluorescence intensity. A method based on the excitation fluorescence intensity correction coefficient matrix is put forward, which eliminates effectively the influence of the asymmetry of array excitation intensity. Data collection and processing for actual samples are made and imaging results are got. The experimental results show the validity of the processing methods and some unsolved problems in the processing methods are discussed.