生物化学技术

Electron crystallography studies the structure of two-dimensional crystals of membrane proteins or other crystalline arrays. This method has been used to determine the atomic structures of six membrane proteins and tubulin, as well as several other structures at a slightly lower res ...

With the rapid progresses in both instrumentation and computing, it is increasingly straightforward and routine to determine the structures of icosahedral viruses to subnanometer resolutions (6–10 Å) by cryoelectron microscopy and image reconstruction. In this resolution ra ...

The evolvement of preparative methods in structural studies has always been as important as the development of sophisticated equipment. Software development is also a significant part for three-dimensional (3D) structural studies using electron tomography methods (ETMs). Ad ...

Standard immunogold-labeling methods in transmission electron microscopy (TEM) are unable to locate immunogold particles in the depth direction. This inability does not only concern bulky whole mounts, but also sections. A partial solution to the problem is stereo inspection. Howe ...

This chapter covers the conventional methods and considerations for preparing cultured cells for examination in the transmission electron microscope. Techniques for handling cells grown in liquid culture, as well as on substrates such as culture dishes or agar, are discussed. Direc ...

Cryoelectron microscopy of frozen-hydrated specimens is currently the only available technique for determining the “native” three-dimensional ultrastructure of individual examples of organelles and cells. Two techniques are available, stereo pair imaging and electron ...

A transmission electron microscope (TEM) accessory, the energy filter, enables the establishment of a method for elemental microanalysis, the electron energy-loss spectroscopy (EELS). In conventional TEM, unscattered, elastic, and inelastic scattered electrons contribu ...

The protocol of cryoplaning techniques that to examine the distribution of water in living tree stems by cryoscanning electron microscopy have been developed and described. In brief, the procedures are as follows: First, a portion of transpiring stem is frozen in the standing state with liqu ...

This chapter covers conventional methods for preparing biological specimens for examination in the scanning electron microscope (SEM). Techniques for handling cells grown in liquid culture, as well as on substrates such as culture dishes, slide culture chambers or agar, are discuss ...

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semithick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and ...

Static secondary ion mass spectrometry (SSIMS) is a capable of providing detailed atomic and molecular characterization of the surface chemistry of biological and biomedical materials. The technique is particularly suited to the detection and imaging of small molecules such as mem ...

Modern microscopy in plant sciences has evolved in the direction of providing ultra-structural and analytical information simultaneously. Energy-dispersive X-ray microanalysis (EDX) is a powerful technique that allows the qualitative and quantitative measurement of ma ...

Biological tissues are passed through numerous procedures before they can be studied at the ultrastructural level with the electron microscope. Chemical fixation is widely used as a method for preserving structural detail and can be performed by simple immersion or total body vascular ...

This chapter describes conventional chemical fixation methods and techniques for studying the cellular and organelle ultrastructure of plant tissues under transmission electron microscopy. The general methods and procedures for the plant specimen preparation (includ ...

This chapter describes the practical details involved in the successful sectioning of plastic-embedded specimens for electron microscopy. The focus will be on those parts of the process that are really key to the successful sectioning of any biological material, i.e., the proper shape and s ...

Microwave processors can provide a means of rapid processing and resin embedding for biological specimens that are to be sectioned and examined by transmission electron microscopy. This chapter describes a microwave-assisted protocol for processing, dehydrating, and embeddi ...

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. In this chapter, techniques are p ...

Lipid-associated disorders are a worldwide health concern and major efforts are directed toward understanding—at the molecular levels—mechanisms of lipid storage and degradation, in healthy and diseased states. Yeast is a widely used model organisms to study such processes at the c ...

Many experiments require isolation and purification of membranes and organelles from a cell-free lysate. A combination of differential and sucrose density gradient centrifugation provides adequate separation of most yeast organelles in a single experiment. Yeast cells are co ...

Visualization of protein–protein interactions in vivo offers a powerful tool to resolve spatial and temporal aspects of cellular functions. Bimolecular fluorescence complementation (BiFC) makes use of nonfluorescent fragments of green fluorescent protein or its variants ...