stained Protocols

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A sensitive method for the detection of apoptosis by single laser flow cytometry. Methodology includes: Staining for detection of apoptosis, Direct Staining Procedure, Indirect Staining Procedure, Protocol for the use of actinomycin D (AD) on samples that were stained with 7-AAD for apoptosis and fixed in formaldehyde. - [Read Apoptosis Detection Protocol By Single Laser Flow Cytometry]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol uses specific antibodies coupled to one of four fluorochromes: fluorescein isothiocyanate (FITC), R-phycoerythrin (PE), peridinin chlorophyll-a (PcP), and allophycocyanin (APC). These fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Category: Immunology: B Cell Protocols

Fluorochromes can be used simultaneously to stain and analyze the expression patterns of four different proteins in the same sample. The fluorochrome stained cell populations are analyzed using a FACSCalibur dual-laser flow cytometer. - [Read Characterization of Cells by Flow Cytometry Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact non-ovule materials of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse root, seedling even flower development without sectioning. This protocol could also be used for clearing GUS stained material, after chlorophyll is removed by 70% ethanol. - [Read Clearing Arabidopsis Non-Ovule Materials With the HCG Solution Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols: Stains for Senile Plaques Protocols

Protocol to demonstrate amyloid deposits in tissue sections. When stained with the Congo Red Stain the amyloid, with the aide of polarizing lenses, will birefringe an apple green color.  Under the microscope. - [Read Congo Red Putcher's Modification Amyloid Protocol]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes how to dissect spinal cords of dsRNA-treated avian embryos for analysis of commissural axon guidance.  After fixation of the spinal cord with paraformaldehyde, commissural axons are stained with Fast-DiI. - [Read Dissection of Spinal Cords for Analysis of Axonal Pathfinding in dsRNA Treated Avian Embryos]

Category: Avian Bird Protocols

Protocol describes how to dissect spinal cords of dsRNA-treated avian embryos for analysis of commissural axon guidance. After fixation of the spinal cord with paraformaldehyde, commissural axons are stained with Fast-DiI. - [Read Dissection of Spinal Cords for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

DNA laddering can be detected from samples with only 8% apoptotic cells. Alternatively, the cells can be stained with DAPI and analysed by flow cytometry. CellDeath.de - [Read DNA laddering assay for treated cells]

Category: Microscopy Protocols

FM 4-64 is a lipophilic styryl dye and a vital stain: it fluoresces only in living cells, so cells cannot be fixed then stained nor stained then fixed. You must stain and observe living cells. FM 4-64 does not permeate cell membranes but, instead, intercalates into the plasma membrane is then taken into the cells by endocytosis. - [Read FM 4-64 Labeling of Yeast Vacuole Membranes Protocol]

Category: Molecular Imaging: Electron Microscopy Protocols

In brief, the reaction mixture, is prepared in exactly the same way as if the sample is to be stained or shadowed for electron microscopy, but once the... Gold labeling technique for electron microscopic identification and location of proteins. - [Read Gold labeling of proteins for electron microscopy]

Category: Neuroscience Protocols: Neuronal Stains Protocols

An appropriate term for glial fibers is 'nerve glue', because they provide the internal support of the central nervous system.  There are four types of glial cells: astrocytes, oligosendroglia,microglia, and ependymal cells. The glia fibers are stained with crystal violet which are resistant to the aniline-chloroform differentiating solution. - [Read Holzer's Stain Protocol]

Category: Proteomic Protocols

In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels. (EMBL protocol). - [Read In Gel Digestion Protocols for Silver Stained and Coomassie Stained Gels]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: DNA Staining Protocols for Flow Cytometry

This protocol describes a method for quantitative measurement of DNA using propidium iodide (PI) staining and flow cytometry. PI stains all double-stranded regions of both DNA and RNA by intercalating between the stacked bases of the double helix. PI cannot penetrate an intact cell membrane; therefore, cells are fixed prior to staining. The ethanol-fixed cells can be stored unstained at 4°C for days, or even weeks, and then stained and analyzed. - [Read Measurement of DNA Content Using Propidium Iodide (PI) Staining of Fixed Whole Cells Protocol]

Category: Drosophila Protocols: Drosophila Staining Protocols

Early embryos (0-17 hours or until cuticle formation) are treated with a mixture of organic solvents, formaldehyde, and alcohols, as described here. The cuticles of late-stage embryos are usually opened by sonication. Tissues from more advanced stages of development are normally dissected by hand and then fixed and stained in a standard paraformaldehyde/detergent combination - [Read Preparing Early Whole-Mount Drosophila Embryos for Immunostaining Protocol]

Category: Drosophila Protocols: Drosophila Staining Protocols

Early and late embryos are treated with a mixture of organic solvents, formaldehyde, and alcohols. The cuticles of late-stage embryos (17-22 hours or until hatching) are usually opened by sonication, as described here. Tissues from later stages of development are normally dissected by hand and then fixed and stained in a standard paraformaldehyde/detergent combination. - [Read Preparing Late Whole-Mount Drosophila Embryos for Immunostaining Protocol]

Category: Immunology: B Cell Protocols

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]

Category: Staining Protocols: Simple Staining Protocols

Protocol for simple staining. Smear is stained with a solution of a single dye which stains all cells the same color. Differentiation of cell types or structures is not the objective of the simple stain. However, certain structures which are not stained by this method may be easily seen, for example, endospores and lipid inclusions. - [Read Simple Staining Protocol]

Category: Primary Cell Isolation and Culture Protocols

Slide Preparation for Manual Microdissection for Subsequent DNA, RNA, and Protein Analysis. Manual microdissection and subsequent molecular analysis can be carried out on slides stained using standard hematoxylin and eosin methods. However, if cell types that are (or are not) expressing a specific protein are required for a study, then more advanced slide preparation methods such as Immuno-LCM may be utilized. - [Read Slide Preparation for Manual Microdissection Protocol]