Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

Spinning disk TIRF microscopy can be a useful tool for studying the role of protein during cytoskeletal network formation. This technique allowed three-dimensional imaging of fast cellular approaches that occur in the cell and at the same the precise localization of fluorescence molecule that are placed at the plasma membrane. This method can provide insight to research areas like microbiology, cell biology, and all those branches where it's important to study the interaction between the plasma membrane and the cellular environment.

This method can be extended to those live imaging experiments where it's important to localize the structures in the plasma membrane and where you also want to maintain a high resolution of the remaining cellular volume. The crucial things one has to consider are the choice of the proper cell line to set up the TIRF illumination and the other imaging parameters and to find the focus of the transfected cells. The visual demonstration of this protocol would certainly help to avoid problems with the handling of the cells during image acquisition.