Live imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows following dynamic cellular processes as they unfold "in vivo", and often reveals a degree of complexity impossible to study in still images. Thanks to the increasing interest in this technique, significant technical improvements have been made in the recent years in microscopy leading to more sensitive and faster cameras, more efficient filter sets and a variety of different microscope systems to choose from. The temporal dimension of cellular processes in microscopy is still often neglected due to the difficulties in technical setup and the necessity to fabricate your own tools. Now the majority of the required equipment is commercially available from microscope manufacturers and related companies including environmental chambers, CO2-control and temperature control.
Often used in conjunction with time-lapse microscopy are fluorescent proteins, such as the Green Fluorescent Protein (GFP) or the Red Fluorescent Protein (RFP; Shaner et al., 2004; Shaner et al., 2005; Wiedenmann et al., 2004) and live dyes. Likewise this is a rapidly developing field of research, which has provided a variety of different "flavors" of fluorescent proteins for the scientific community, with properties specifically suited for different live-imaging applications (for a guide to choose the right one, see Shaner et al., 2005).
MPI of Immunobiology
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