embryos Protocols

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Aggregation of ES cells and eight cell stage embryos. Bowtell Lab. - [Read Aggregation of ES cells and eight cell stage embryos]

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: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Aggregation of ES Cells with Mouse Morula-state Embryos. Oxford Practical Series Approach. - [Read Aggregation of ES Cells with Mouse Morula-state Embryos]

Category: Cell Biology and Cell Culture

Protocol applies EFs to cells in vitro but has been modified and to use electrotactic chambers to accommodate cells growing in planar culture or in three-dimensional (3D) gels, en bloc tissue cultures in 3D and possible small embryos, such as that from frog and zebra fish. The EF is applied to the cells or tissues cultured in a customer designed electrotactic chamber via agar salt bridges, Steinberg’s solution and Ag/AgCl electrodes. - [Read Application of Direct Current Electric Fields to Cells and Tissues in vitro]

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 diploid embryos. If embryos from a heterozygous mutant intercross are aggregated with wild-type embryos, the resulting chimeras can be used for analyzing mutant phenotypes. - [Read Assembling Aggregates between Diploid 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: Xenopus Protocols

Protocol describes the use of a basic water-based dye to stain for acid mucosubstances and acetic mucins located on the cartilage of fixed embryos, permitting the examination of cartilage formation. - [Read Cartilage Staining of Xenopus tropicalis Protocol]

Category: Stem Cell Protocols: Stem Cell Injection Protocols

This protocol describes a method for collecting two-cell- to compacted morula-stage embryos. Such embryos can then be used for microinjection. - [Read Collecting Two-Cell- to Compacted Morula-Stage Embryos Protocol]

Category: RNAi Technology Protocols: RNAi Mouse Embryos and Oocytes Protocols

Protocol describes a method to collect early embryos from 6-week-old mice.  Subsequently, the isolated embryos can be injected with double-stranded RNA to induce knockdown of a gene of interest. - [Read Collection of Early Mouse Embryos for RNAi Protocol]

Category: Transgenic Mouse Protocols: Mouse Genotyping Protocols

Practical Approach Online, Oxford University Press. Combined LacZ staining and in situ hybridization of mouse embryos, beta-galactosidase. - [Read Combined LacZ staining and in situ hybridization PDF]

Category: RNAi Technology Protocols: RNAi Drosophila Protocols

Protocol describes how subcellular-sized particles are accelerated to high velocity to carry double-stranded RNA (dsRNA) into Drosophila embryos.  The major advantage of this procedure over microinjection (Microinjection of dsRNA into Drosophila Embryos) is that particle bombardment is easier and faster to perform.  In addition, the mechanical trauma received is far less than by microinjection, allowing better survival of embryos and fewer phenotypic artifacts. - [Read Delivery of dsRNA into Drosophila Embryos by Gene Gun Protocol]

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol for detection of even-skipped transcripts in Drosophila embryos with PCR/DIG-labeled DNA probes. - [Read Detection of Even-Skipped Transcripts in Drosophila Embryos with PCR/DIG-Labeled DNA Probes]

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for detection of even-skipped transcripts in drosophila embryos with PCR/DIG-labeled DNA probes. This protocol has been used to detect the transcript distribution of a number of genes by in situ hybridization, including evenskipped and seven-up, in whole mount Drosophila embryos, and engrailed Antennapedia in whole mount grasshopper embryos. Includes: Probe labeling; Evaluation of labeling reaction; Preparation of embryos, hybridization and detection.
 - [Read Detection of Even-Skipped Transcripts in Drosophila Embryos with PCR/DIG-Labeled DNA Probes Protocol]

Category: Xenopus Protocols: Xenopus Injection Protocols

Differences in injection of X. laevis and X. tropicalis. Includes: X. tropicalis lays eggs about 4 hours after a boost of hCG; In vitro fertilization is not nearly as efficient in trops compared to laevis; X. tropicalis embryos are much softer than X. laevis embryos; X. tropicalis embryos whose jelly coats are removed by cysteine have a loose sticky vitelline membrane; X. tropicalis do not have a "summer slump". - [Read Differences in Injection of X. laevis and X. tropicalis]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for disaggregating embryos and the ICM of blastocysts into individual cells. - [Read Disaggregating Cleavage-Stage Embryos and the Inner Cell Mass of Blastocysts into Individual Cells]

Category: Xenopus Protocols

Information on the diseases of Xenopus tropicalis. Includes: PREMETAMORPHS: Bacterial Infection of Embryos; Protozoal Infestation of Tadpoles; POSTMETAMORPHS: Mycobacteriosis; Chytridiomycosis (Batrachochytrium dendrobatidis). - [Read Diseases of Xenopus tropicalis]

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: C. elegans Protocols: C. elegans Fixing Protocols

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

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

ES Cell Injection Into Mouse Embryos. Practical Approach Online, Oxford University Press. - [Read ES Cell Injection Into Mouse Embryos]

Category: Developmental Biology: Embryology Protocols

The FAM caspase binding assay kits from ATCC Corporation can be used to determine amounts of active caspases in cells. The FAM-labeled caspase inhibitor can freely diffuse into the cell. Active caspase irreversibly binds the inhibitor. Upon washing the cells, the amount of fluorescence is proportional to the amount of active caspase in the cell. FAM-LETD-fmk (catalog no. 30-1306) is used to detect caspase 8 and FAM-LEHD-fmk (catalog no. 30-1308) is used for caspase 9. - [Read Fam Caspase 8 and 9 Binding Assay for Embryos 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: Mouse Protocols: Mouse Embryology Protocols

Protocol provides methods for fixation of mouse embryos and tissues with paraformaldehyde, Bouin’s fixative, or methanol/DMSO solution. - [Read Fixation of Mouse Embryos and Tissues Protocol]