FISH and immunocytochemistry: towards visualising single target molecules in living cells.
Knowledge of how molecules interact in space and time is crucial for understanding cellular processes. A host of novel techniques have been developed for the visualisation of single target molecules in living cells, many based on fluorescence in situ hybridisation (FISH) or immunocytochemistry (IC). To extend the applicability of FISH to living cells, special backbone-modified probes and specific conformations (molecular beacons) have been designed. In the case of IC, conventional immunoreagents have been fine-tuned with respect to size and affinity or replaced with new protein scaffolds based on ankyrin repeat proteins. Other key advances include the use of proximity ligation to confirm vicinity binding and the use of quantum dots, which have proven potential for cellular labelling.
Tanke
Hans J
HJ
Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands. h.j.tanke@lumc.nl
Dirks
Roeland W
RW
Raap
Ton
T
D002468:Cell Physiological Phenomena; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008967:Molecular Biology / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004247:DNA