Contemporary biology has developed into the era of cell biology and molecular biology, and people try to study the
mechanism of all kinds of biological phenomena at the microcosmic level now. Accurate description of the
microstructure of biological samples is exigent need from many biomedical experiments. This paper introduces a method
for 3-dimensional quantitative observation on the microstructure of vital biological samples based on two photon laser
scanning microscopy (TPLSM). TPLSM is a novel kind of fluorescence microscopy, which has excellence in its low
optical damage, high resolution, deep penetration depth and suitability for 3-dimensional (3D) imaging. Fluorescent
stained samples were observed by TPLSM, and afterward the original shapes of them were obtained through 3D image
reconstruction. The spatial distribution of all objects in samples as well as their volumes could be derived by image
segmentation and mathematic calculation. Thus the 3-dimensionally and quantitatively depicted microstructure of the
samples was finally derived. We applied this method to quantitative analysis of the spatial distribution of chromosomes
in meiotic mouse oocytes at metaphase, and wonderful results came out last.
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