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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 3 of 3.

Publications

Eschen-Lippold, L.; Landgraf, R.; Smolka, U.; Schulze, S.; Heilmann, M.; Heilmann, I.; Hause, G.; Rosahl, S. Activation of defense against Phytophthora infestans in potato by down-regulation of syntaxin gene expression New Phytol 193, 985-996, (2012) DOI: 10.1111/j.1469-8137.2011.04024.x

The oomycete Phytophthora infestans is the causal agent of late blight, the most devastating disease of potato. The importance of vesicle fusion processes and callose deposition for defense of potato against Phytophthora infestans was analyzed.Transgenic plants were generated, which express RNA interference constructs targeted against plasma membrane‐localized SYNTAXIN‐RELATED 1 (StSYR1) and SOLUBLE N‐ETHYLMALEIMIDE‐SENSITIVE FACTOR ADAPTOR PROTEIN 33 (StSNAP33), the potato homologs of Arabidopsis AtSYP121 and AtSNAP33, respectively.Phenotypically, transgenic plants grew normally, but showed spontaneous necrosis and chlorosis formation at later stages. In response to infection with Phytophthora infestans, increased resistance of StSYR1‐RNAi plants, but not StSNAP33‐RNAi plants, was observed. This increased resistance correlated with the constitutive accumulation of salicylic acid and PR1 transcripts. Aberrant callose deposition in Phytophthora infestans‐infected StSYR1‐RNAi plants coincided with decreased papilla formation at penetration sites. Resistance against the necrotrophic fungus Botrytis cinerea was not significantly altered. Infiltration experiments with bacterial solutions of Agrobacterium tumefaciens and Escherichia coli revealed a hypersensitive phenotype of both types of RNAi lines.The enhanced defense status and the reduced growth of Phytophthora infestans on StSYR1‐RNAi plants suggest an involvement of syntaxins in secretory defense responses of potato and, in particular, in the formation of callose‐containing papillae.
Publications

Zdyb, A.; Demchenko, K.; Heumann, J.; Mrosk, C.; Grzeganek, P.; Göbel, C.; Feussner, I.; Pawlowski, K.; Hause, B. Jasmonate biosynthesis in legume and actinorhizal nodules New Phytol 189, 568-579 , (2011) DOI: 10.1111/j.1469-8137.2010.03504.x

Jasmonic acid (JA) is a plant signalling compound that has been implicated in the regulation of mutualistic symbioses. In order to understand the spatial distribution of JA biosynthetic capacity in nodules of two actinorhizal species, Casaurina glauca and Datisca glomerata, and one legume, Medicago truncatula, we determined the localization of allene oxide cyclase (AOC) which catalyses a committed step in JA biosynthesis. In all nodule types analysed, AOC was detected exclusively in uninfected cells.The levels of JA were compared in the roots and nodules of the three plant species. The nodules and noninoculated roots of the two actinorhizal species, and the root systems of M. truncatula, noninoculated or nodulated with wild‐type Sinorhizobium meliloti or with mutants unable to fix nitrogen, did not show significant differences in JA levels. However, JA levels in all plant organs examined increased significantly on mechanical disturbance.To study whether JA played a regulatory role in the nodules of M. truncatula, composite plants containing roots expressing an MtAOC1‐sense or MtAOC1‐RNAi construct were inoculated with S. meliloti. Neither an increase nor reduction in AOC levels resulted in altered nodule formation.These data suggest that jasmonates are not involved in the development and function of root nodules.
Publications

Mielke, K.; Forner, S.; Kramell, R.; Conrad, U.; Hause, B. Cell-specific visualization of jasmonates in wounded tomato and Arabidopsis leaves using jasmonate-specific antibodies New Phytol 190, 1069-1080, (2011) DOI: 10.1111/j.1469-8137.2010.03638.x

Jasmonates are well‐characterized signals in the development of plants and their response to abiotic and biotic stresses, such as touch and wounding by herbivores. A gap in our knowledge on jasmonate‐induced processes, however, is the cellular localization of jasmonates.Here, a novel antibody‐based approach was developed to visualize jasmonates in cross‐sections of plant material. Antibodies raised in rabbits against BSA‐coupled jasmonic acid (JA) are specific for JA, its methyl ester and isoleucine conjugate. They do not bind to 12‐oxophytodienoic acid, 12‐hydoxy‐JA or coronatine. These antibodies were used in combination with newly established fixation and embedding methods.Jasmonates were rapidly and uniformly distributed within all cells near the site of damage of a mechanically wounded tomato (Solanum lycopersicum) leaf. Leaf tissue distally located to the wound site exhibited identical distribution, but had a lower signal intensity. The occurrence of jasmonates in all cell types of a wounded leaf was accompanied by transcript accumulation of early JA‐induced genes visualized by in situ hybridization.With these new antibodies, a powerful tool is available to detect cell‐specifically the occurrence of jasmonates in any jasmonate‐dependent stress response or developmental process of plants.

This page was last modified on 14.11.2018.

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