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丁香实验推荐阅读
Assessment of Sphingosine-1-Phosphate Receptor Expression and Associated Intracellular Signaling Cascades in Primary Cells of the Human Central Nervou

Measuring the effects of sphingosine-1-phosphate (S1P) receptor modulators on human primary neural cells is of particular interest given the recent application of these central nervous system-accessible agents to the treatment of neurodegenerative diseases, such as multip ...

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Use of Intravital Microscopy and In Vitro Chemotaxis Assays to Study the Roles of Sphingosine-1-Phosphate in Bone Homeostasis

We describe a method to visualize the migration of osteoclast precursors within intact murine bone �marrow in real time using intravital multiphoton microscopy. Conventionally, cell migration has been evaluated using in vitro systems, such as transmigration assays. Although the ...

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Production of Tissue-Engineered Skin and Oral Mucosa for Clinical and Experimental Use

Since the early 1990s, our understanding of how epithelial and stromal cells interact in 3D tissue-engineered constructs has led to tissue-engineered skin and oral mucosa models, which are beginning to deliver benefit in the clinic (usually in small-scale reconstructive surgery pro ...

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Genetic Modification of Chondrocytes Using Viral Vectors

The use of isolated cells to construct engineered tissues provides the opportunity to genetically modify those cells prior to the formation of tissue. This should make it possible to create transgenic human model tissues that can be used to determine gene function as well as to identify or valid ...

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Stem Cell and Neuron Co-cultures for the Study of Nerve Regeneration

Many experimental in vivo studies have indicated that Schwann cells are key facilitators of peripheral nerve regeneration but their clinical therapeutic potential may be limited. Recent advances suggest that stem cell therapy could one day be used to treat nerve traumas. We have shown how a ...

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Quantification of mRNA Using Real-Time PCR and Western Blot Analysis of MAPK Events in Chondrocyte/Agarose Constructs

In vitro models of chondrocyte mechanobiology have been used to compare the intracellular signalling pathways altered in normal and osteoarthritis-affected cartilage. However, differences in the model system and type of loading configuration have led to complicated pathways. ...

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Analytical Technologies for Integrated Single-Cell Analysis of Human Immune Responses

The immune system is a network of cells in which the constitutive members interact through dense and sometimes overlapping connections. The extreme complexity of this network poses a significant challenge for monitoring pathological conditions (e.g., food allergies, autoimmun ...

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Introduction: Why Analyze Single Cells

Powerful methods in molecular biology are abundant; however, in many fields including hematology, stem cell biology, tissue engineering, and cancer biology, data from tools and assays that analyze the average signals from many cells may not yield the desired result because the cells of inte ...

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Identification of Enzyme-Converted Peptide Products from Single Cells Using Capillary Electrophoresis and Liquid Chromatography-Mass Spectrometry

Single-cell analysis using chemical methods, otherwise known as chemical cytometry, promises to provide significant leaps in understanding signaling processes which result in cellular behavior. Sensitive methods for chemical cytometry such as capillary electrophores ...

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Flow Cytometry and Microscopy as Means of Studying Single Cells: A Short Introductional Overview

Flow cytometry and microscopy are perhaps the two most commonly used techniques for analyzing single cells. Both techniques are typically robust and provide a high throughput analysis of living and/or fixed cells. The techniques are often combined with fluorescent labeling of cells, us ...

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Laser Capture Microdissection for Gene Expression Analysis of Specific Cell Populations in Single Blastocysts

Laser capture microdissection (LCM) allows for the isolation of small tissue fractions from heterogeneous tissue sections, for downstream genetic or proteomic analysis without contamination by the surrounding tissue. This technique can also be successfully used for the isola ...

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Droplet Microfluidics for Single-Cell Analysis

This book chapter aims at providing an overview of all the aspects and procedures needed to develop a droplet-based workflow for single-cell analysis (see Fig. 10.1). The surfactant system used to stabilize droplets is a critical component of droplet microfluidics; its properties define t ...

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Screening of Antigen-Specific Antibody-Secreting Cells

Screening of antigen-specific antibody-producing cells is a key step for obtaining antigen-specific monoclonal antibodies. In murine system, hybridoma between B-lymphocytes and myeloma cells is used to screen and produce antigen-specific monoclonal antibodies. In human sy ...

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Analysis of Single Eukaryotic Cells Using Raman Tweezers

Raman Tweezers is a technique that combines optical trapping with Raman spectroscopy and has enabled the spectroscopic analysis of single cells. Applications of this technique include the identification and discrimination of different types of cells, including healthy and non- ...

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Single-Cell Microinjection Technologies

Single-cell microinjection, as a mechanical delivery tool, has been used for transferring substances into transfection or infection challenging cells. Here, we discuss the advantages and applications of microinjection, list the materials needed for performing microinject ...

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Ultrasonic Manipulation of Single Cells

Ultrasonic manipulation has emerged as a simple and powerful tool for trapping, aggregation, and separation of cells. During the last decade, an increasing amount of applications in the microscale format has been demonstrated, of which the most important is acoustophoresis (continu ...

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Single-Cell Culture in Microwells

In order to better understand cellular processes and behavior, a controlled way of studying high numbers of single cells and their clone formation is greatly needed. This chapter describes a microwell plate with 672 wells in a standard array/slide format, applied for single-cell culture and ...

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Isodielectric Separation and Analysis of Cells

Measuring the electrical properties of a cell provides a fast and accessible means of identifying or characterizing cells whose biological state differs from the population as a whole. This chapter describes a microfluidic method for characterizing the electrical properties of ce ...

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Single Cell Electroporation Using Microfluidic Devices

Electroporation is a powerful technique to increase the permeability of cell membranes and subsequently introduce foreign materials into cells. Pores are created in the cell membrane upon application of an electric field (kV/cm). Most applications employ bulk electroporation, at ...

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Perfusion Culture of Mammalian Cells in a Microfluidic Channel with a Built-In Pillar Array

In vitro culture of mammalian cells is fundamental to various biological studies such as single cell analysis, pathological research, and drug/therapy development. However, there are limitations with the current in vitro cell culture methods. Cells tend to lose their specific functi ...

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