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丁香实验推荐阅读
Two-Electrode Voltage Clamp

Two-electrode voltage clamp (TEVC) is a conventional electrophysiological technique used to artificially control the membrane potential (V m) of large cells to study the properties of electrogenic membrane proteins, especially ion channels. It makes use of two intracellular ele ...

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Recording Single-Channel Currents Using Smart Patch-Clamp Technique

Microdomains that form on the plasma membrane of cells are essential for signalling compartmentation within cells. The localization of ion channels in these surface microdomains is important in defining what signalling cascades will be generated. For example, in cardiomyocytes, s ...

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Automated Planar Patch-Clamp

Ion channels are integral membrane proteins that regulate the flow of ions across the plasma membrane and the membranes of intracellular organelles of both excitable and non-excitable cells. Ion channels are vital to a wide variety of biological processes and are prominent components of t ...

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Piezo-Electrically Driven Mechanical Stimulation of Sensory Neurons

Mechanotransduction, the conversion of a mechanical stimulus into a biological response, constitutes the basis of a variety of physiological functions such as the senses of touch, balance, proprioception, blood pressure, and hearing. In vertebrates, mechanosensation is mediat ...

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Perforated Whole-Cell Patch-Clamp Recording

Perforated whole-cell patch-clamp is a variant of the patch-clamp technique used to measure the sum activity of ion channels in the plasma membrane of a single cell. Its defining feature is that electrical access to the cell is obtained through inclusion of a pore-forming antibiotic in the patch p ...

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Combined Single-Channel and Macroscopic Recording Techniques to Analyze Gating Mechanisms of the Large Conductance Ca2+ and Voltage Activated (BK) Pot

Ion channels are integral membrane proteins that regulate membrane potentials and signaling of cells in response to various stimuli. The patch-clamp technique enables the study of single channels or a population of channels. The macroscopic recording approaches are powerful in rev ...

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Viral Gene Delivery: Optimized Protocol for Production of High Titer Lentiviral Vectors

HIV-derived lentiviral vectors (LVV) are among the most commonly used gene delivery vehicles. Their production in high quantities, which enables concentration of viral particles to high titers, is important for their successful application in both biomedical research and gene the ...

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Transient Overexpression of Genes in Neurons Using Nucleofection

Nucleofection is a transfection method used to introduce substrates such as cDNA plasmids into primary cells or other cell lines. The method can be successfully applied to cells that are considered difficult to transfect or suffer from low transfection efficiency as seen with tradition ...

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Use of Escherichia coli for the Production and Purification of Membrane Proteins

Individual types of ion channels and other membrane proteins are typically expressed only at low levels in their native membranes, rendering their isolation by conventional purification techniques difficult. The heterologous over-expression of such proteins is therefore us ...

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Mammalian Expression Systems and Transfection Techniques

To delineate the function of a single ion channel subtype amongst the multitude that normally constitutes a signalling pathway, it is often insightful to study the function or signalling of that one ion channel in the absence of the others. Mammalian cell lines that do not normally express the gene of ...

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Approaches to Cloning of Pain-Related Ion Channel Genes

Molecular pain research is a relatively new and rapidly expanding field that represents advancement in conventional pain research. One of the fundamentals of molecular pain involves the cloning of genes and especially the ion channels specifically involved in nociceptive proces ...

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Quantification of Cyclobutane Pyrimidine Dimers in Human Epidermal Stem Cells

The common procedures that are used to quantify cyclobutane pyrimidine dimers (CPD) comprise the extraction of cellular DNA followed by the detection of this nucleic acid modification by immunoblotting or electrophoretic methods. Consequently, these approaches provide an ave ...

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Monitoring the Cycling Activity of Cultured Human Keratinocytes Using a CFSE-Based Dye Tracking Approach

The development of methods and tools suitable for functional analysis of keratinocytes placed in an in vitro context is of great importance for characterizing properties associated with their normal state, for detecting abnormalities related to pathological states, or for studyi ...

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Identification and Analysis of Epidermal Stem Cells from Primary Mouse Keratinocytes

The skin, one of the largest organs of the body, is a dynamic tissue in which terminally differentiated keratinocytes are replaced by the proliferation and differentiation of epidermal stem cells. Epidermal stem cells are relatively undifferentiated, retain a high capacity for self-r ...

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Isolation and Characterization of Cutaneous Epithelial Stem Cells

During homeostasis, adult mammalian skin turnover is maintained by a number of multipotent and �unipotent epithelial progenitors located either in the epidermis, hair follicle, or sebaceous gland. Recent work has illustrated that these various progenitor populations reside in r ...

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Lineage Tracing of Hair Follicle Stem Cells in Epidermal Whole Mounts

Lineage tracing of tissue stem cells represents a powerful tool to address fundamental questions of deve�lopment, differentiation and cellular renewal in a natural tissue environment. The Cre/lox site-specific recombination system is increasingly used to genetically label s ...

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Analysis of Bulge Stem Cells from the Epidermis Using Flow Cytometry

The epidermis is a multilayered epithelium consisting of multiple different progenitor cell populations, all of which are important to epidermal function. In order to study these populations, several techniques have been developed that enable specific purification of the diffe ...

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Isolation of Hair Follicle Bulge Stem Cells from YFP-Expressing Reporter Mice

In this article we provide a method to isolate hair follicle stem cells that have undergone targeted gene inactivation. The mice from which these cells are isolated are bred into a Rosa26-yellow fluorescent protein (YFP) reporter background, which results in YFP expression in the targeted st ...

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Isolation and Cultivation of Human Scalp Interfollicular Epidermal Stem Cells

Skin regeneration is intricately controlled by epidermal stem cells. In human skin, the long-lived, slow-cycling, and highly proliferative stem cells are located in the basal layer of the interfollicular epidermis (IFE). The ability to isolate and culture human IFE stem cells (IFESCs) of ...

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Skin Stem Cells

The skin stem cells are located at two distinct locations, the basement of epidermis and the hair follicle bulge. The bulge stem cell is considered to be of higher hierarchy in terms of the stemness than the epidermal stem cell. Recently, hair follicle bulge cells can be successfully isolated using fl ...

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