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Technique Advances and Applications of Cryo-Electron-Microscopy for the Study of Vitrified Biological Systems- [electronic resource]

자료유형: 학위논문파일 국외

최종처리일시: 20240214101215

ISBN: 9798380314879

DDC: 530

저자명: Yu, Yue.

서명/저자: Technique Advances and Applications of Cryo-Electron-Microscopy for the Study of Vitrified Biological Systems - [electronic resource]

발행사항: [S.l.]: : Cornell University., 2023

발행사항: Ann Arbor : : ProQuest Dissertations & Theses,, 2023

형태사항: 1 online resource(117 p.)

주기사항: Source: Dissertations Abstracts International, Volume: 85-03, Section: B.

주기사항: Advisor: Kourkoutis, Lena.

학위논문주기: Thesis (Ph.D.)--Cornell University, 2023.

사용제한주기: This item must not be sold to any third party vendors.

초록/해제: 요약Cryo-electron microscopy (cryo-EM) is a powerful technique extensively utilized for studying biological systems. It allows for high-resolution structural determination of macromolecules and visualization of cellular structures in their near-native environment. This thesis starts with a concise overview of the fundamental principles and applications of selective cryo-EM techniques with conventional transmission electron microscopy (TEM), such as single particle analysis (cryo-SPA), electron tomography (cryo-ET), and focused-ion-beam milling (cryo-FIB). Subsequently, applications of scanning-TEM (STEM) in vitrified hydrated biological systems are introduced, along with the progress made in four-dimensional STEM (4DSTEM) techniques.The challenges encountered in imaging thick biological specimens using conventional TEM techniques are highlighted, along with the limited applicability of conventional STEM for high-resolution structural determination due to collection efficiency limitations. To address these challenges, a 4D-STEM technique, tilt-corrected bright-field STEM (tcBF-STEM) is introduced, aiming to provide advantages for imaging thick dose-sensitive specimens by optimizing the collection efficiency of coherent BF-STEM images. The implementation of tcBF-STEM with a pixelated detector optimized for STEM is demonstrated, along with the extraction of probe aberration information using tcBF imaging. Notably, tcBF-STEM enables the retrieval of information beyond the real-space scanning Nyquist limit, effectively mitigating the effects of specimen motion in cryo-EM experiments with a single tilt direction. The results of applying tcBFSTEM to imaging vitrified biological specimens, including vitrified apoferritin protein and E. coli bacteria, are presented. The potential of tcBF-STEM and its limitations are discussed at the end.