3D confocal reconstruction of gene expression in mouse.
Three-dimensional computer reconstructions of gene expression data will become a valuable tool in biomedical research in the near future. However, at present the process of converting in situ expression data into 3D models is a highly specialized and time-consuming procedure. Here we present a method which allows rapid reconstruction of whole-mount in situ data from mouse embryos. Mid-gestation embryos were stained with the alkaline phosphotase substrate Fast Red, which can be detected using confocal laser scanning microscopy (CLSM), and cut into 70 microm sections. Each section was then scanned and digitally reconstructed. Using this method it took two days to section, digitize and reconstruct the full expression pattern of Shh in an E9.5 embryo (a 3D model of this embryo can be seen at genex.hgu.mrc.ac.uk). Additionally we demonstrate that this technique allows gene expression to be studied at the single cell level in intact tissue.
Hecksher-Sørensen
J
J
Department of Comperative and Developmental Genetics, MRC Human Genetics Unit, Western General Hospital, Crewe Road, EH4 2XU, UK.
Sharpe
J
J
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D004396:Coloring Agents / Q000494:pharmacology; D004064:Digestive System / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism*; D015870:Gene Expression; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods*; D008565:Membrane Proteins / Q000096:biosynthesis; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D009499:Neutral Red / Q000494:pharmacology; D000072079:Patched Receptors; D014176:Protein Biosynthesis; D011956:Receptors, Cell Surface; D012984:Software; D015534:Trans-Activators / Q000096:biosynthesis
D004396:Coloring Agents; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D008565:Membrane Proteins; D000072079:Patched Receptors; D011956:Receptors, Cell Surface; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D009499:Neutral Red; D000469:Alkaline Phosphatase