细胞功能测定

Glioblastoma multiforme (GBM) are the most common malignant brain tumours in adults, characterized by short survival periods of patients. Their aggressive local growth pattern and increased invasiveness, due to a high motility of the tumour cells, hamper treatment. However, the mole ...

Embryonic stem (ES) cells are an important source of stem cells in tissue engineering and regenerative medicine because of their high self-renewal capacities and differentiation potentials. However, the detailed molecular mechanisms controlling the differentiation and ren ...

Single-cell manipulation supporting robot (SMSR) has enabled femtoinjection, a high-throughput and semi-quantitative microinjection in the range of femtogram (fg) DNA and other molecules. An enhanced green fluorescent protein (EGFP) gene expression vector can be introduced ...

Osteoclasts and their precursors have traditionally been considered difficult cells to transfect using standard approaches. Here, we describe several methods for transfection of mature osteoclasts and their precursors using the Amaxa™ Nucleofector system, lentiviruses, ...

Transcription is the process by which the rate of RNA synthesis is regulated. Here, we describe the techniques for carrying out promoter-reporter assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays, three commonly used methods for studying g ...

This chapter describes the analysis of signalling pathways in bone cells by the use of western blotting and immunoprecipitation, including a step by step guide to cell culture techniques, protein isolation, purification, measurement, electrophoretic transfer, and detection.

Here, we present methods for extracting DNA and RNA from samples of whole bone tissue and culture bone cells and describe methods quantitative and qualitative measurement of the extracted nucleic acids. These protocols described provide high-quality nucleic acids suitable for down ...

Bone histomorphometry remains an important tool to study the pathophysiology of bone disease and the cellular mechanism by which treatments work. Here, we review the methods for embedding, sectioning, staining, and analysis of bone sections in rodents.

This chapter describes methods for quantitation of gene expression in bone cells and bone tissues using the technique of quantitative reverse transcription-polymerase chain reaction.

Immunohistochemistry (IHC) is a routinely used technique in clinical diagnosis of pathological conditions and in basic research. It combines anatomical, immunological, and biochemical methods and relies on the specific binding of an antibody to an antigen. Using the technique with ...

Here, we described a method for carrying out nonradioactive in situ hybridization to detect mRNA transcripts in cryostat sections of mouse bone using the CryoJane� Tape-Transfer System and digoxigenin (DIG)-labeled riboprobes.

There has been great interest in the identifying the mechanisms by which apoptosis is regulated in bone over recent years and in the biological role that this process plays in bone metabolism and bone disease. Here, we describe several methods for the detection of apoptosis in bone sections and in bo ...

This chapter described methods for Scanning Electron Microscopical imaging of bone and bone cells. Backscattered electron (BSE) imaging is by far the most useful in the bone field, followed by secondary electrons (SE) and the energy dispersive X-ray (EDX) analytical modes. This chapter co ...

This chapter describes procedures to process mineralized tissues obtained from different sources for transmission electron microscopy (TEM). Methods for fixation, resin embedding, staining of semi-thin sections and ultrathin sections are presented. In addition, attenti ...

Over the past two decades there have been unprecedented advances in the capabilities for live cell imaging using light and confocal microscopy. Together with the discovery of green fluorescent protein and its derivatives and the development of a vast array of fluorescent imaging probes a ...

In order to understand osteoclast cell biology, it is necessary to culture these cells on a physiological �substrate that they can resorb in vitro, such as bone or dentine. However, this creates problems for analysis by fluorescence microscopy, due to the depth of the sample under investigation. ...

This chapter describes the use of digital micro-radiography with the Faxitron machine as a means of imaging and quantitating bone mineral content in mice and rats.

This chapter provides information for the use of peripheral quantitative computed tomography in small animals, including suggestions for study design, instrument setting, and data interpretation.

This chapter describes the use of microcomputed tomography scanning for analysing bone structure, focussing on rodent bone. It also discusses sample preparation, the correct set-up of the scanner, and the impact of some of the important scanner settings.

Optical imaging is a valuable technique for visualizing and quantifying biological processes in living �organisms. Optical imaging can be divided into two main imaging modalities: bioluminescence imaging and fluorescence imaging. This chapter describes the use of these imagi ...