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丁香实验推荐阅读
Viability Assessment of Osteocytes Using Histological Lactate Dehydrogenase Activity Staining on Human Cancellous Bone Sections

The assessment of viable osteocytes within bone tissue is of crucial importance. Osteocytes are the most abundant cells in bone. Due to their interconnectivity in the bone matrix they are hypothesised to play an important role in the maintenance of the extracellular matrix of bone. The death of o ...

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Confocal Imaging Protocols for Live/Dead Staining in Three-Dimensional Carriers

In tissue engineering, a variety of methods are commonly used to evaluate survival of cells inside tissues or three-dimensional (3D) carriers. Among these methods confocal laser scanning microscopy opened accessibility of 3D tissue using live cell imaging into the tissue or 3D scaffold ...

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Fluorescein Diacetate for Determination of Cell Viability in 3D FibroblastCollagenGAG Constructs

Quantification of cell viability and distribution within engineered tissues currently relies on representative histology, phenotypic assays, and destructive assays of viability. To evaluate uniformity of cell density throughout 3D collagen scaffolds prior to in vivo use, a no ...

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Cytotoxicity Testing: Measuring Viable Cells, Dead Cells, and Detecting Mechanism of Cell Death

Testing the effects of compounds on the viability of cells grown in culture is widely used as a predictor of potential toxic effects in whole animals. Among the several alternative assays available, measuring the levels of ATP is the most sensitive, reliable, and convenient method for monitori ...

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Multiplexing Cell Viability Assays

Today, obtaining mechanistic insights into biological, toxicological, and pathological processes is of upmost importance. Researchers aim to obtain as many as possible data from one cell sample to understand the biological processes under study. Multiplexing, which is the ability ...

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Rapid Quantification of Cell Viability and Apoptosis in B-Cell Lymphoma Cultures Using Cyanine SYTO Probes

The gross majority of classical apoptotic hallmarks can be rapidly examined by multiparameter flow cytometry. As a result, cytometry became a technology of choice in diverse studies of cellular demise. In this context, a novel class of substituted unsymmetrical cyanine SYTO probes has re ...

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Measurement of Caspase Activity: From Cell Populations to Individual Cells

Caspases are critical regulators of the apoptotic program, responsible for the harmonic dismantling of the cell. Cell death can occur by way of different options (necroptosis, necrosis, extreme autophagy) but once caspases are fully engaged it will take the apoptotic route. Hence, in gene ...

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Annexin V/7-AAD Staining in Keratinocytes

Annexin V/7-amino-actinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Early apoptotic cells express phosphatidylserines (PS) on the outer leaflet of the plasma membrane. PS can be stained by labeled annexin V. Late apoptotic cel ...

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Analysis of Tumor and Endothelial Cell Viability and Survival Using Sulforhodamine B and Clonogenic Assays

A variety of assays, and rationales for their use, exist to monitor viability and/or survival following cellular exposure to insult. Two commonly used in vitro assays are the sulforhodamine B assay and the clonogenic survival assay which can be used to monitor the efficacy of anticancer agents, ...

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The xCELLigence System for Real-Time and Label-Free Monitoring of Cell Viability

We describe here the use of the xCELLigence system for label-free and real-time monitoring of cell �viability. The xCELLigence system uses specially designed microtiter plates containing interdigitated gold microelectrodes to noninvasively monitor the viability of cultur ...

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Assessment of Cell Proliferation with Resazurin-Based Fluorescent Dye

The Alamar Blue assay is based on enzymatic reduction of indicator dye by viable cells and serves as an effective tool for assessing cell proliferation and as a screening technique. It can be applied in studies concentrating on animal, plant, yeast, and bacteria cells. Among the various methods for ...

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WST-8 Analysis of Cell Viability During Osteogenesis of Human Mesenchymal Stem Cells

WST-8 is one of the newer generation formazan-based dyes, which release the converted product into the medium in a soluble form. This allows for a non-destructive determination of viability enabling the cells to be subject to further investigations. This is a major advantage in cases where cell p ...

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Estimation of Cell Number Based on Metabolic Activity: The MTT Reduction Assay

The MTT reduction assay is used to determine the level of metabolic activity in eukaryotic cells, �including animal, plant, and fungal cells. If the metabolic rate is constant, the technique can be employed to count living cells in a sample. Once it is set up, the method is very robust, and can be automatized to ...

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Cell Viability Analysis Using Trypan Blue: Manual and Automated Methods

One of the traditional methods of cell viability analysis is the use of trypan blue dye exclusion staining. This technique has been the standard methodology used in academic research laboratories and industrial biotechnology plants. Cells were routinely counted manually with a hemoc ...

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Digital Image Processing of Live/Dead Staining

The quantification of live and dead cells in a substrate is often an essential step in cell biology research. A staining protocol that acts differently on live and on dead cells is applied and the number of cells visible is counted using a microscope. Often this counting is done manually or only evaluated ...

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Using Endogenous MicroRNA Expression Patterns to Visualize Neural Differentiation of Human Pluripotent Stem Cells

Many existing protocols for neuronal differentiation of human pluripotent cells result in heterogeneous cell populations and unsynchronized differentiation, necessitating the development of methods for labeling specific cell populations. Here we describe how microR ...

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Single-Cell Transcript Profiling of Differentiating Embryonic Stem Cells

Heterogeneity of stem cell populations is a well-known but poorly characterized phenomenon. Here, we demonstrate the qualitative and quantitative power of single-cell transcript analysis to characterize transcriptome dynamics in embryonic stem cells (ESC). In this chapter, we ...

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Neural Differentiation of Human ES and iPS Cells in Three-Dimensional Collagen and Martigel Gels

In this chapter, we describe an effective and reproducible protocol for neural differentiation of human pluripotent stem cells in three dimensional (3D) collagen and MartigelTM gels. We have used this protocol to generate embryoid bodies (EBs) from dissociated suspension cultures of ...

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Direct Differentiation of Human Embryonic Stem Cells into Selective Neurons on Nanoscale Ridge/Groove Pattern Arrays

Human embryonic stem cells (hESCs) are pluripotent cells that have the potential to be used for tissue engineering and regenerative medicine. Biochemical and biological agents are widely used to induce hESC differentiation. However, it would be better if we could induce the differentia ...

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Generation of Lung Epithelial-Like Tissue from hESC by AirLiquid Interface Culture

Human embryonic stem cells (hESCs) have the capacity to differentiate in vivo and in vitro into cells from all three germ lineages. The in vitro generation of lung cells and tissues from hESCs creates opportunities for fundamental research, drug development or cell-replacement therapy. In ...

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