Protocol 1: Microdissection of sucrose treated brain tissues
Isolation of specific subanatomical regions or single cells in the central nervous system is a delicate procedure requiring careful preparation of tissue specimen. In areas with low cell density singe cells may be collected, while in areas with high density of cells within a cellular layer (e.g. granular cell layers of the cerebellum, dentate gyrus, or CA1-3 areas of the hippocampus) clusters consisting of the same cell types may be collected with each single acquisition step†. Separation of cell bodies and dendrites can also be performed in certain areas of the brain, such as the hippocampus and cerebellum. This protocol describes the isolation of single cells, cell clusters, and subanatomical regions from sucrose treated brain slices. Sample microdissections are provided to demonstrate the effectiveness of this method.
Protocol 2: Microdissection of native brain tissue for neural progenitor cultures
Our instrument is an efficient and effective way for the isolation of subanatomical regions from live brain tissue slices. The vacuum pulls the cells into the capillary with minimal mechanical damage, and large areas can be collected quickly using disposable capillaries (DCU) with greater inner diameters. This protocol describes the technique established to isolate living cells from live brain sections for primary neural progenitor cell culture. A simple protocol for the collection of various subanatomical brain regions and the successful subsequent recultivation of stem cell cultures is described here.
Protocol 3: Collection and clonal expansion of individual cells from adherent cultures
This application note is aimed at demonstrating the performance of CIU on adherent cell cultures and determined that this is a suitable method for individual cell collection and subsequent clonal expansion. the instrument also accurately and rapidly acquired both fluorescently labeled and non-labeled individual cells directly from standard culture dishes, and more importantly, the collected cells were suitable for subsequent recultivation.
Protocol 4: Microdissection of heart muscle and other hard to dissect tissues
This protocol is suitable for the dissection of muscles and other difficult to dissect tissues and isolation of high quality RNA from microdissected material.