METHOD AND APPARATUS FOR DISCRIMINATING X AND Y SPERMS BASED UPON RAMAN SPECTROSCOPY

摘要

The invention relates to a method based upon the use of Raman spectroscopy to face the problem of the biochemical analysis and of the differentiation of the X sperm and Y sperm in the animal species, in particular bovine species, with approaches which make possible the subsequent use in fecundation techniques. A method addressed to the resolution of this problem is based upon the separation between X sperm and Y sperm before insemination. An apparatus is also described implementing such method which allows to distinguish and characterize the difference between X sperm and Y sperm by acquiring and analyzing Raman spectra noninvasively and in a not destructive way. By means of a statistical approach, such as for example the analysis of the main components and the linear discriminating analysis, the spectroscopic difference between X sperm and Y sperm is evaluated, processed and used to separate the two types of cells. The invention also relates to the development of a cytofluorimeter combining the Raman microscope with a system for trapping optically the sperms and a microfluidic chamber. More precisely, the optical trapping system, equipped with an objective with high numerical aperture, is used to immobilize the cells and, the same objective is used to excite and gather Raman spectra. At last, the microfluidic chamber is used to make the sample to flow, with the final purpose of providing a quick, precise and not invasive separation of X sperm and Y sperm, (figure 5A).