Establishing Yeast 3D Image and Increasing Its Images Definition with Focused Ion Beam Dual Beam Microscope
Date Issued
2010
Date
2010
Author(s)
Chen, Yu
Abstract
In recent years, the technology of the electron microscope grows vigorously, not only make the research of the biological field have considerable progress, but develop in the instrument and increase with knowledge give us more understanding of life and relation of the microcosms. In this thesis, we utilized yeast as a target. Focused ion beam dual beam microscope is different from the scanning electron microscope and confocal microscope. It could not only offer the information on a surface of samples, but no need to prepare the sample into a flake. Namely, it can get sectional information which were cut by ion beam to observe the micro-structure within samples, and then can assemble those series pictures to three-dimensional images of the whole cell. It is a quite powerful tool to provide whole cells'' information for studying morphology. The traditional chemical treatment is complicated and usually accompanied with poisonous medicines, after the process of fixation, dehydration and embedding, the slight structure of biological cells often might run off or deform in the course, and different from real situation. The tool we used, Focused ion beam microscope (FIB) Nova 600i, carries cryo-system, because the biological sample was frozen rapidly by liquid nitrogen; the structures that could be observed should most close to truth. We could observe the micro-structures in the section images of the frozen fresh yeast. While making three-dimensional image to rebuild whole cell, the factor for judging the secondary structure is very important, so we have made a succession of experiments to compare the difference of 3D images between fresh cell and traditional treated cell and to contrast their section images. Understand information and its advantage that cryo system and FIB can offer in the research of the biological structure.
Subjects
Focused ion beam
FIB
dual beam
yeast
3D
reconstruction
Type
thesis
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