The purpose of this project is to map the locations at which specific genes are expressed in pre-implantation mouse embryos. Dr. Minoru Ko's group in LG has used expression profiling with microarrays to identify 500-1000 genes that are differentially expressed at various stages of early mouse embryo development. Initially, fluorescence in-situ hybridization (FISH) will be used to probe the locations within the embryo where these genes are expressed. Three dimensional restoration microscopy (the DeltaVision system from Applied Precision, Inc) will be employed to image these fluorescent probes. In conjunction with other fluorescent cellular markers for DNA, cytoskeletal components, and membranes, we plan to build a 3-dimensional model of the early mouse embryo in a standardized orientation, so that we can compare the location and level of gene expression from different imaging experiments. Eventually a database will be constructed to link the imaging experiments to microarray data and other mouse genomics data sources. We have found two probe technologies that show promise for this application: 3-DNA and quantom dots. Recently, we have made significant progress in increasing specificity of the probe and reducing fluorescence background
| Yoshikawa, Toshiyuki; Piao, Yulan; Zhong, Jinhui et al. (2006) High-throughput screen for genes predominantly expressed in the ICM of mouse blastocysts by whole mount in situ hybridization. Gene Expr Patterns 6:213-24 |
