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丁香实验推荐阅读
Application of SEM and EDX in Studying Biomineralization in Plant Tissues

This chapter describes protocols using formalin-acetic acid-alcohol (FAA) to fix plant tissues for studying biomineralization by means of scanning electron microscopy (SEM) and qualitative energy-dispersive X-ray microanalysis (EDX). Specimen preparation protocols f ...

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Three-Dimensional Imaging of Adherent Cells using FIB/SEM and STEM

In this chapter we describe three different approaches for three-dimensional imaging of electron microscopic samples: serial sectioning transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM) tomography, and focused ion beam/scann ...

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Correlative 3D Imaging: CLSM and FIB-SEM Tomography Using High-Pressure Frozen, Freeze-Substituted Biological Samples

Correlative light and electron microscopy aims at combining data from different imaging modalities, ideally from the same area of the one sample, in order to achieve a more holistic view of the hierarchical structural organization of cells and tissues. Modern 3D imaging techniques opened ...

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Correlative Light and Electron Microscopy Using Immunolabeled Sections

In correlative microscopy, light microscopy provides the overview and orientation of the complex cells and tissue, while electron microscopy offers the detailed localization and correlation of subcellular structures. In this chapter we offer detailed high-quality electron m ...

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FIB-SEM Tomography in Biology

Three-dimensional information is much easier to understand than a set of two-dimensional images. Therefore a layman is thrilled by the pseudo-3D image taken in a scanning electron microscope (SEM) while, when seeing a transmission electron micrograph, his imagination is challenged. F ...

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Correlative Fluorescence and Electron Microscopy of Quantum Dot Labeled Proteins on Whole Cells in Liquid

Correlative fluorescence microscopy and scanning transmission electron microscopy (STEM) of cells fully immersed in liquid is a new methodology with many application areas. Proteins, in live cells immobilized on microchips, are labeled with fluorescent quantum dot (QD) nanopa ...

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Nanometer-Resolution Fluorescence Electron Microscopy (Nano-EM) in Cultured Cells

Nano-resolution fluorescence electron microscopy (nano-fEM) pinpoints the location of individual proteins in electron micrographs. Plastic sections are first imaged using a super-resolution fluorescence microscope and then imaged on an electron microscope. The two imag ...

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Correlative Light and Electron Microscopy: From Live Cell Dynamic to 3D Ultrastructure

Correlative light and electron microscopy (CLEM) aims at combining data acquired from the same sample through both imaging modalities. Many combinations can be found in the literature where almost any kind of light microscopy (LM) has been associated to different processing in electron ...

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Electron Tomography for Organelles, Cells, and Tissues

Electron tomography (ET) is an emerging electron microscopy (EM) technique for three-dimensional (3D) visualization of molecular arrangements and ultrastructural architectures in organelles, cells, and tissues at 2–10 nm resolution. The 3D tomogram is reconstructed from a ser ...

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Single Particle Cryo-electron Microscopy and 3-D Reconstruction of Viruses

With fast progresses in instrumentation, image processing algorithms, and computational resources, single particle electron cryo-microscopy (cryo-EM) 3-D reconstruction of icosahedral viruses has now reached near-atomic resolutions (3–4 �). With comparable resoluti ...

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Biological Applications of Phase-Contrast Electron Microscopy

Here, I review the principles and applications of phase-contrast electron microscopy using phase plates. First, I develop the principle of phase contrast based on a minimal model of microscopy, introducing a double Fourier-transform process to mathematically formulate the image fo ...

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Visualization of DNA and ProteinDNA Complexes with Atomic Force Microscopy

This article describes sample preparation techniques for AFM imaging of DNA and protein–DNA complexes. The approach is based on chemical functionalization of the mica surface with aminopropyl silatrane (APS) to yield an APS-mica surface. This surface binds nucleic acids and nucleop ...

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Tracking DNA and RNA Sequences at High Resolution

In this chapter, the basic principles and protocols of the electron microscopical detections of specific DNA and RNA sequences are described. We focused primarily on a comparison of various methods of electron microscopy in situ hybridization (EM-ISH) with respect to their sensitivity ...

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Cryo-electron Microscopy of Membrane Proteins

Electron crystallography is used to study membrane proteins in the form of planar, two-dimensional (2D) crystals, or other crystalline arrays such as tubular crystals. This method has been used to determine the atomic resolution structures of bacteriorhodopsin, tubulin, aquapori ...

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Analysis of Specificity in Immunoelectron Microscopy

Immunoelectron microscopy (immuno-EM) using gold labeling on sections is a powerful technique for mapping the distribution of proteins, lipids, carbohydrates, and nucleic acids in intact biological systems. The gold particles comprise a useful and readily quantifiable digital ...

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Electron Microscopy of Microtubule Cytoskeleton Assembly In Vitro

Cell-free cytoplasm isolated from meiotic Xenopus egg extracts reconstitutes microtubule phenomena in vitro. These crude extracts assemble bipolar meiotic spindles and are readily fractionated for biochemical assays, providing a good tool to dissect molecular mechanism. We ...

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Negative Staining and Cryo-negative Staining: Applications in Biology and Medicine

Negative staining is widely applicable to isolated viruses, protein molecules, macromolecular assemblies and fibrils, subcellular membrane fractions, liposomes and artificial membranes, synthetic DNA arrays, and also to polymer solutions and a variety of nanotechnology s ...

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Cryo-electron Microscopy of Vitreous Sections

More than 30 years ago two groups independently reported the vitrification of pure water, which was until then regarded as impossible without a cryoprotectant . This opened the opportunity to cryo-electron microscopy (cryo-EM) to observe biological samples at nanometer scale, close to ...

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In Vitro and In Vivo Demonstrations of Fluorescence by Unbound Excitation from Luminescence (FUEL)

Bioluminescence imaging is a powerful technique that allows for deep-tissue analysis in living, intact organisms. However, in vivo optical imaging is compounded by difficulties due to light scattering and absorption. While light scattering is relatively difficult to overcome and ...

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Noninvasive In Vivo Monitoring of Extracellular Vesicles

Extracellular vesicles (EVs) including exosomes and microvesicles are nanometer-sized vesicles released by cells to deliver lipids, cellular proteins, mRNAs, and noncoding RNAs, thereby facilitating intercellular communication without direct cell-to-cell contact ...

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