For most of our imaging, we use a Zeiss LSM 880 confocal microscope. Our unit is equipped with Airyscan and produces super resolution images of fluorescently labeled proteins in a variety of cell lines.
We are interested in the colocalization of proteins to the nucleus as well as during the cell cycle. We also induce DNA damage to observe if further proteins interactions are apparent. Multiple proteins are tagged with primary and secondary antibodies to observe what role they play in the cell.
Confocal image of HeLa cells stained with MAb 414. MAb414 is an excellent rim stainer and provides a clear distinction between regions inside and outside of the nucleus.
Confocal image of DT40 cells tagged with Ran and Lamin B. Colocalization is apparent in the merged image since the red and green channel overlay to produce yellow. Furthermore, the linear pattern of the scatterplot indicates that pixels from the red and green channels overlap and that they are actually correlated.
Live cell imaging of DT40 cells with nuclear (blue) and endoplasmic reticulum (green) staining.
Live cell imaging is another capability of the LSM 880. So far, we have used stably transfected cells containing GFP-tagged proteins to study their localization under live cell conditions.
Microscope image of Galdieria sulphuraria.
Credit: Gerald Schoenknecht