High resolution analysis of mitotic metaphase chromosomes with scanning electron microscopy [Elektronische Ressource] : localizing histone H3 modifications with immunogold labeling in barley (Hordeum vulgare) / submitted by Elizabeth Schroeder-Reiter

High resolution analysis of mitotic metaphase chromosomes with scanning electron microscopy:Localizing histone H3 modifications with immunogold labeling in barley (Hordeum vulgare)Dissertationfrom theDepartment of Biology IElectron MicroscopyLudwig-Maximillians-Universität MünchenSubmitted byElizabeth Schroeder-ReiterAugust 20041. Referee: Prof. Dr. G. Wanner2. Referee: Prof. Dr. P. DittrichDate of oral defense: 22 October, 20042Table of ContentsAbbreviations ................................................................................4Introduction...................................................................................6Materials and Methods.................................................................9Peparation of plant material ..........................................................................9Enzymatic tissue dissociation .......................................................................10Laser marked slides ......................................................................................10Drop/Cryo fixation and isolation technique for chromosomes.....................10Alternative isolation technique with ”suspension prparation”....................11Enzymatic treatment for removal of nucleoplasm.........................................11DNA staining.................................................................................................12Immunolabeling ...................................................