embryo Protocols

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Protocol descibes the use of L929 mouse fibroblast cells cultured in vitro in an agarose overlay assay to assess the toxicity of test substances.  The assay may be useful in assessing the irritation potential of test substances (e.g. surfactant-based products) as an alternative to the Draize rabbit eye test. - [Read Agarose Overlay Assay Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the aggregation of ES cells with mouse morula-stage embryos. Includes: Preparation of aggregation walls; ES cell preparation; Embryo preparation and aggregation. - [Read Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: Plant Biology Protocols

The Allium test provides a rapid screening procedure for chemicals, pollutants contaminants, etc. which may represent environmental hazards.  Root growth inhibition and adverse effects upon chromosomes provide an indication of likely toxicity. - [Read Allium Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Microorganism Cytotoxicity Assays Protocols

The AMES procedure evaluates the mutagenic potential of test chemicals by their effect on five histidine requiring strains of the bacterium, Salmonella typhimurium in the absence and presence of a rat liver metabolising system. - [Read Ames Test]

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing diploid embryo-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for diploid embryo-diploid embryo aggregation. The resulting chimeras are useful for phenotypic analysis when an induced mutation has an extraembryonic phenotype. - [Read Assembling Aggregates between Diploid and Tetraploid Embryos Protocol]

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for assembling aggregates between ES cells and diploid embryos. The resulting chimeras are useful for separating certain extraembryonic phenotypes from phenotypes in the embryo proper, since the diploid embryo contributes to all parts of the conceptus, but the ES cell component does not contribute to the trophoblast or yolk sac endoderm. - [Read Assembling Aggregates between Embryonic Stem (ES) Cells and Diploid Embryos Protocol]

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing ES cell-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for ES cell-diploid embryo aggregation in which diploid embryos are replaced with tetraploid embryos. The resulting chimeras can be used to analyze the embryonic versus extraembryonic phenotype of a mutation. - [Read Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos Protocol]

Category: Radioactivity: Autoradiography

Antonio Simeone. MRC Centre for Developmental Neurobiology, New Hunt’s Houre. - [Read Autoradiography for mRNA detection in. mouse embryo tissue sections. PDF]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol for blastocyst embryo transfer into pseudopregnant recipient female mouse. - [Read Blastocyst Embryo Transfer into Pseudopregnant Mouse Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

Bovine spermatozoa cytotoxicity test. - [Read Bovine Spermatozoa Cytotoxicity Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Human A431 cells and mouse 3T3 cells are exposed in culture to UV light both in the presence and absence of test compound.  Phototoxicity is expressed as a decrease in cell viability as determined by the MTT assay. - [Read Cell Culture Phototoxicity Test Protocol]

Category: Plant Biology Protocols

Protocol can be used for clearing intact seeds of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse embryo and endosperm development without sectioning. - [Read Clearing Arabidopsis Ovule Materials with the Hoyer's Solution Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The potential cytotoxicity of compounds under hypoxic conditions is determined by exposing cell cultures to test compounds in a low oxygen atmosphere.  Subsequent cell survival is determined by the MTT and methylene blue colorimetric assays. - [Read Colorimetric Cytotoxcity Assays for Anchorage Dependent Cells Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

This bioassay utilizes cultured Hepa-lclc7 (Hepa-1) mouse hepatoma cells to assess the CYPlA1-inducing potency or cytotoxicity of pure test chemicals or environmental samples.  In the Hepa-l induction test , the CYPlA1-inducing potency of the test sample is detected as increased aryl hydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase (EROD) activities. - [Read CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Changes in the balance of cytoskeletal proteins after exposure to test compounds can be detected by indirect immunofluorescence microscopy and quantitative biochemical methods. - [Read Cytoskeletal Alterations as a Parameter for Assessment of Toxicity]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

WiCell Research Institute. Recipes, Frequently Asked Questions, Mouse Embryo Extraction and Tissue, Harvesting Mouse Embryonic Fibroblasts (MEF)from mice. - [Read Derivation of Mouse Embryonic Fibroblasts (MEF) Part I PDF]

Category: Drosophila Protocols

Protocol for Drosophila embryo microinjection. - [Read Drosophila Embryo Microinjection Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Macrophage cells in culture may be exposed to particulate matter, and resultant effects upon cell viability determined by vital dye exclusion and enzyme leakage assays. - [Read Dust Toxicity in Rat Alveolar Macrophage Cultures Protocol]

Category: C. elegans Protocols: C. elegans Fixing Protocols

Protocol for EM fixation of C. elegans embryos. - [Read EM Fixation of C. elegans Embryos Protocol]

Category: Xenopus Protocols: Xenopus Embryo Manipulation Protocols

Protocol for embryo lysates and immunoprecipitation: protein A agarose & protein A magnetic beads. Includes: Embryo lysates; Immunoprecipitation with protein A agarose; Immunoprecipitation with protein A magnetic beads for silver stain/sequencing analysis. - [Read Embryo Lysates & Immunoprecipitation Protocol]

Category: Drosophila Protocols: Drosophila Genetics Protocols

FISH protocols for Drosophila. Includes: RNA Probe Preparation; Embryo Collection and Fixation; Single FISH on Drosophila embryos; Post-Fixation, Hybridization and Post-Hybridization Washes; Development of FISH Signal; Storage, Mounting and Viewing of Samples; Double FISH on Drosophila Embryos; RNA-Protein Double Labeling; FISH on Dissected Tissues. - [Read FISH Protocols for Drosophila]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

Protocol describes a method for performing isoelectric fractionation of a maize embryo sample using a multicompartment electrolyzer(MCE). This prefractionation of proteins having pIs within a certain pH interval is essential for allowing high loads of protein to be resolved on narrow and ultra-narrow immobilized pH gradients used in 2D electrophoresis. The isoelectric membranes in the MCE act like isoelectric traps capturing all the protein species having pIs encompassing the pI value of each... - [Read Fractionation of Maize Embryo Proteins for 2-D Gel Electrophoresis Using Multicompartment Electrolyz]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The cytotoxic effect of chemicals upon cells in culture is measured by cell viability (neutral red uptake) method. - [Read Frame Modified Neutral Red Uptake Cytotoxcity Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

This bioassay utilises cultured H-4-II-E rat hepatoma cells to assess the aryl hydrocarbon hydroxylase (AHH) inducing potencies of planar aromatic hydrocarbons and/or contaminated environmental samples.  The response of the cells to pure test chemicals or extracts of mixtures is compared with their response to the standard 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). - [Read H-4-II-E Rat Hepatoma Cell Bioassay Protocol]