title: Suitable transfection methods for single particle tracing in plant suspension cells.
abstract: BACKGROUND
  A multitude of different imaging systems are already available to image genetically altered RNA species; however, only a few of these techniques are actually suitable to visualize endogenous RNA. One possibility is to use fluorescently-labelled and hybridization-sensitive probes. In order to yield more information about the exact localization and movement of a single RNA molecule, it is necessary to image such probes with highly sensitive microscope setups. More challenges arise if such experiments are conducted in plant cells due to their high autofluorescence and demanding transfection procedures.
  
  
  RESULTS
  Here, we report in planta imaging of single RNA molecules using fluorescently labeled molecular beacons. We tested three different transfection protocols in order to identify optimal conditions for transfection of fluorescent DNA probes and their subsequent detection at the single molecule level.
  
  
  CONCLUSIONS
  We found that an optimized heat shock protocol provided a vastly improved transfection method for small DNA molecules which were used for subsequent single RNA molecule detection in living plant suspension cells.
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohrgasse 9/3, Vienna, Austria.
authors.forename: Janett
authors.initials: J
authors.lastname: Göhring
authors.affiliation: Gregor Mendel Institute, Dr. Bohrgasse 3, Vienna, Austria.
authors.forename: Nick
authors.initials: N
authors.lastname: Fulcher
authors.affiliation: Upper Austria University of Applied Sciences, Campus Linz, Linz, 4020, Austria.
authors.forename: Kurt
authors.initials: K
authors.lastname: Schilcher
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohrgasse 9/3, Vienna, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Barta
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohrgasse 9/3, Vienna, Austria ; Upper Austria University of Applied Sciences, Campus Linz, Linz, 4020, Austria.
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Jacak