化学生物学实验技术

Two methods for the cryopreservation of mammalian oocytes are described. One method uses a relatively low concentration of the cryoprotectant propanediol plus sucrose and requires controlled-rate cooling equipment to achieve a slow cooling rate. Such a method has produced live bir ...

The cryopreservation of mammalian embryos has expanded over the past 20 yr by encompassing a range of sophisticated methods to deal with different developmental stages and different sensitivities to low-temperature exposure. We have described a method for slow, controlled-rate fr ...

This chapter provides an up-to-date overview of freeze-drying (lyophilization) with particulars relevance to stabilizing live cells or viruses for industrial applications as vaccines or seed culture. The chapter discusses the importance of formulation, cycle development, v ...

Cryopreservation is the use of very low temperatures to preserve structurally intact living cells and tissues. Unprotected freezing is normally lethal and this chapter seeks to analyze some of the mechanisms involved and to show how cooling can be used to produce stable conditions that pre ...

This chapter describes the methods that can be applied to successfully freeze-dry proteins. Laboratory applications are given at small scale, typified by the purification of a protein intermediate as part of the analytical characterization of a protein, and at intermediate scale, as il ...

Traditionally, the ex situ study of prokaryotes (members of Archaea and Bacteria) has required access to living cultures. Although it may be possible to maintain a small number of strains by serial transfer, such methods are not the best, particularly for long-term storage or for keeping a large co ...

A method is described that allows a wide range of yeast species to be stored in liquid nitrogen while maintaining a high level of viability. Yeast cultures are sealed in commercially available polypropylene straws after having been mixed with a glycerol-based cryoprotectant. Once placed in a ...

A method is described that allows yeast species to be stored using a variation on the standard freeze-drying method, which employs evaporative cooling in a two-stage process. Yeast cultures are placed in glass ampoules after having been mixed with a lyoprotectant. Primary drying is carried o ...

Recent technical innovations in mass spectrometry-based techniques have resulted in a range of highly sensitive and versatile instruments for high-throughput, high-sensitive, proteome-scale profiling. This wide diversity of instrumentation commercially available ...

Within ecosystems microorganisms coexist and interact. Knowledge of these interactions is of great importance in the fields of ecology, food production, and medicine. Such interactions often involve the synthesis of antibiotic secondary metabolites. Different kinds of s molec ...

T lymphocytes, including cytotoxic CD8+ T cells, are important cells involved in immunology, as they can destroy infected or tumor cells. We describe here a detailed protocol starting from CD8+ T lymphocytes isolation for T cell culture followed by total protein extraction or subcellular fr ...

An understanding of gene function requires a complementation of gene and gene expression analysis by the systematic analysis of proteins. Progress in plant proteomics has been lagging behind animal and microbial proteomics due to the lack of plant genome data and the problems involved in s ...

In this chapter we describe a method to analyze human serum with the goal of discovering disease-related changes in the serum proteome. The methodology is based on the removal of the six most abundant serum proteins by immunoaffinity chromatography. This step is followed by trypsin digestion ...

Cell membrane microparticles (MPs) are phospholipid microvesicles shed from the plasma membrane of most eukaryotic cells undergoing activation or apoptosis. The presence of MPs is common in healthy individuals. However, an increase in their release is a controlled event and is consid ...

Identification of binary protein-protein interactions is a crucial step in determining the molecular context and functional pathways of proteins. State-of-the-art proteomics techniques provide high-throughput information on the content of proteomes and protein comple ...

Miniaturized instrumentation and reactors have attracted great interest in the last decade. The first reported use of a microchip was in 1979, when a gas chromatograph air analyzer was fabricated on a silicon wafer (1). It was not until several years later, when flow injection analysis was perfo ...

The advent of photolithography literally brought about the integrated circuit (IC) revolution of the latter part of the twentieth century. Almost all electronic devices that we use today have one or more ICs inside. Improving lithography techniques led to smaller and smaller transisto ...

The lift-off technique is one of the most prevalent methods for fabricating microelectrodes on a flat surface (e.g., a silicon wafer). It represents an alternative for metaletching techniques that often utilize hazardous chemicals in order to define a pattern. This chapter presents an exa ...

We detail the widely prevalent technique of polydimethylsiloxane (PDMS) molding using SU-8 for creating microfluidic chambers and channels. Although other techniques such as injection molding are more apt for mass manufacturing and cost-effective, PDMS molding is used almost ex ...

The use of microfluidic devices is making rapid inroads in the modern laboratory. Traditionally, devices have been manufactured in silica owing to its well-understood surface chemistry and micromachining techniques that are ubiquitous in the microelectronics industry. Recen ...