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Imaging Unit

Nowadays, the elucidation of molecular and biochemical processes requires its combination with investigation of cells – their physiological properties, their structure, the organelles they contain, interactions with their environment, their life cycle, division and death. For centuries, progress in biological research has been connected to the development of tools and equipment that allow new insights into living matter. The invention of and improvements in optical systems were very important because exceeding the limits of the optical resolution of the human eye delivered new insights into tissues, cells and subcellular compartments on the one hand and cellular processes on the other.

The Imaging Unit of the IPB aims to support all research groups in their use of cell biological methods. Currently, at least 13 research groups of the institute are using this unit.

The unit provides:

  • Coordinated supervision and maintenance of equipment
  • Optimal training of coworkers
  • Maximal use of IPB investments
  • Updating of equipment according to state-of-the-art and to requirements of actual research

The working principle of this unit is:

  • Unit headed by one scientist (Prof. Dr. Bettina Hause) and supported by one technician (Hagen Stellmach)
  • Equipment is located decentralized, but its maintenance is carried out centralized
  • For all cell biological methods advice, training and help is provided, extensive experiments, however, have to be done by the co-workers themselves

Devices and materials:

 

Microscopes:

Several stereo microscopes
(Zeiss and Nikon)

Multipurpose MacroMicroSystem equipped with epi-fluorescence: AZ100 (Nikon) with camera (one in each Dept. MSV and SEB)

Lightsheet Microscope
 

Lightsheet Z1 (Zeiss)                                                                                                                                               

Epi-fluorescence microscopes

Axioplan 2 (Zeiss) with differential interference contrast (DIC) device and ApoTome to obtain optical sections, with two cameras (AxioCam MRm and AxioCam MRc5)

AxioImager (Zeiss) with differential interference contrast (DIC) device and ApoTome to obtain optical sections, with two cameras (AxioCam MRm and AxioCam MRc5)

Confocal Laser Scanning Microscope

LSM780 (Zeiss) with Airyscan

LSM700 (Zeiss)


Microtomes

Rotary microtomes to perform semi-thin sections (Microm und Leica)

Vibrating microtome (Vibratome VT1000S, Leica) (Dept. SZB)

Cryo-Microtom CM1950 (Leica)                                                                      

Miscellaneous

InsituPro VSi (Intavis) for automated in situ detection (Dept. SZB)

Micromanipulator (Eppendorf)

Laser Capture Microdissection                                                                    

More devices

  • Organelle-marker: Vectors and transgenic lines of Arabidopsis (Nelson et al., 2007)
  • Wave-marker: Vectors and transgenic lines of Arabidopsis (Geldner et al., 2009)









Methods established:

  • Fixiation, embedding and sectioning of plant materials
  • Laser-Micro-Dissection
  • Immuno labelling
  • in situ-hybridisation
  • light microscopy including fluorescence microscopy
  • confocal laser scanning microscopy
  • Determination of protein interactions via FRET and BiFC (Split-YFP)

Publications by Tag: Cell Biology

Displaying results 1 to 1 of 1.

Publications

Stenzel, I.; Otto, M.; Delker, C.; Kirmse, N.; Schmidt, D.; Miersch, O.; Hause, B.; Wasternack, C. ALLENE OXIDE CYCLASE (AOC) gene family members of Arabidopsis thaliana: tissue- and organ-specific promoter activities and in vivo heteromerization J Exp Bot 63, 6125-6138, (2012) DOI: 10.1093/jxb/ers261

Jasmonates are important signals in plant stress responses and plant development. An essential step in the biosynthesis of jasmonic acid (JA) is catalysed by ALLENE OXIDE CYCLASE (AOC) which establishes the naturally occurring enantiomeric structure of jasmonates. In Arabidopsis thaliana, four genes encode four functional AOC polypeptides (AOC1, AOC2, AOC3, and AOC4) raising the question of functional redundancy or diversification. Analysis of transcript accumulation revealed an organ-specific expression pattern, whereas detailed inspection of transgenic lines expressing the GUS reporter gene under the control of individual AOC promoters showed partially redundant promoter activities during development: (i) In fully developed leaves, promoter activities of AOC1, AOC2, and AOC3 appeared throughout all leaf tissue, but AOC4 promoter activity was vascular bundle-specific; (ii) only AOC3 and AOC4 showed promoter activities in roots; and (iii) partially specific promoter activities were found for AOC1 and AOC4 in flower development. In situ hybridization of flower stalks confirmed the GUS activity data. Characterization of single and double AOC loss-of-function mutants further corroborates the hypothesis of functional redundancies among individual AOCs due to a lack of phenotypes indicative of JA deficiency (e.g. male sterility). To elucidate whether redundant AOC expression might contribute to regulation on AOC activity level, protein interaction studies using bimolecular fluorescence complementation (BiFC) were performed and showed that all AOCs can interact among each other. The data suggest a putative regulatory mechanism of temporal and spatial fine-tuning in JA formation by differential expression and via possible heteromerization of the four AOCs.

This page was last modified on 14.11.2018.

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