实验仪器使用技术

Growth cone collapse is an easy and efficient test for detecting and characterizing axon guidance activities secreted or expressed by cells. It can also be used to dissect signaling pathways by axon growth inhibitors and to isolate therapeutic compounds that promote axon regeneration. H ...

Luciferase reporter systems are widely employed to provide a quantitative readout of gene expression for studies of transcriptional regulation, translation efficiency, and cell signaling. The most common application of luciferase involves transient transfections into c ...

Protein–DNA interactions are critical to maintain genome stability, DNA replication, chromosome �segregation and to regulate gene expression. Chromatin immunoprecipitation (ChIP) is a powerful technique to study these interactions within living neurons and nervous tis ...

The lacZ gene product, β-galactosidase, has classically been used as a reporter of gene expression. β-Galactosidase activity can be detected using a chromogenic substrate, X-gal, which leaves an intense blue precipitate when cleaved by the enzyme. Insertion of the lacZ coding DNA targeted ...

Immunofluorescence (IF), a form of immunohistochemistry (IHC) with specific applications, is commonly used for both basic research and clinical studies, including diagnostics, and involves visualizing the cellular distribution of target molecules (e.g., proteins, DNA, and sm ...

In situ hybridization (ISH) is a type of hybridization that uses a labeled complementary DNA or RNA strand (i.e., probe) to localize a specific DNA or RNA sequence in a portion or section of tissue (In Situ) or in the entire tissue (whole mount ISH). Localization of endogenous transcripts is a desirable app ...

In utero electroporation has been extensively used to study a variety of developmental questions in the developing brain. This protocol aims to provide the basic knowledge for a beginner to get familiar with the technique. Basically, by electroporating a DNA construct into a subpopulati ...

In ovo electroporation is a popular technique to study gene function during development. This technique enables precise temporal and spatial genetic manipulation with the added advantages of being quick and inexpensive. In this chapter the transient transfection of a construct into ...

The developing spinal cord is a well-established model system widely used to study the signaling pathways and genetic programs that control neuronal/glial differentiation and neural circuit assembly. This is largely due to the relatively simple organization (compared to other CNS ...

Lentiviral vectors have become very useful tools for transgene delivery. Based on their ability to transduce both dividing and nondividing cells and to produce long-term transgene expression, lentiviruses have found numerous applications in the biomedical sciences, including ...

Biolistic transfection and diolistic labeling are techniques in which subcellular-sized particles, coated with DNA and lipophilic dyes, respectively, are propelled into cells. The gene-gun approach is particularly applicable for use on ex vivo organized tissue such as brain slic ...

The calcium phosphate transfection is a widely used method for introducing foreign DNA plasmids into cells. Mechanisms underlying this transfection method are not yet defined; however, DNA–calcium phosphate precipitates are internalized by the cells and DNA is efficiently expre ...

Primarily cultured Schwann cells are essential for the investigation of molecular mechanisms regulating proliferation, survival, differentiation, and myelination of Schwann cell and for the development of efficient transplantation for regeneration of injured spinal c ...

The use of enriched oligodendrocyte lineage cell cultures has yielded insight into functions of these cells and regulatory mechanisms. This chapter details methods that result in such cultures.

The use of cultures has informed us of functions and regulation of astrocytes that were previously unknown. This chapter details the methods that result in such cultures.

Dopaminergic neurons are involved in a variety of normal brain functions; degenerations of these neurons cause diseases in human. Investigation of how dopaminergic neurons respond to extracellular signals and molecular mechanisms regulating dopaminergic neuron survival a ...

The cerebellum plays an important role in motor control, motor skill acquisition, memory and learning among other brain functions. In rodents, cerebellar development continues after birth, characterized by the maturation of granule neurons. Cerebellar granule neurons (CGNs) are ...

Nucleoside analog pulse labeling is an important technique which can assess the birthdate, cell cycle maintenance, or cycling rates of cells during development. This method has evolved over several decades of use and is now applied to a multitude of tissue subtypes and systems. The methodolo ...

Pluripotent stem cells are promising potential sources for cell replacement therapy and are useful research tools for exploring disease mechanisms. Neural cells are one of the cell types that have been most efficiently differentiated through several established protocols. This c ...

Neural progenitor cells (NPCs) or neural stem cells are important tools for investigating central nervous system (CNS) development. NPCs can be used in therapeutic strategies and for characterizing differentiation mechanisms. Here, we describe methods for isolating and cultur ...