Labs at Location - Centre of the region Hanà for Biotechnological and Agricultural Research
labs@location Partner
Centre of the Region Haná for Biotechnological and Agricultural Research (CRH)

Department of Cell Biology

Centre of the Region Haná for Biotechnological and Agricultural Research

Centre of the Region Haná for Biotechnological
and Agricultural Research
Department of Cell Biology
Faculty of Science
Palacky University Olomouc
Slechtitelu 11
783 71 Olomouc
Czech Republic
  • Confocal Laser Scanning Microscopy
  • Spinning Disc Microscopy
  • Wide-Field Light Microscopy

Decide on your ideal ZEISS microscope in the facilities of our labs@location partner, the Department of Cell Biology at the Centre of the Region Haná for Biotechnological and Agricultural Research (CRH), Palacký University Olomouc.

The CRH conducts basic and oriented research focused on signal transduction, cytoskeleton and vesicular trafficking during plant development and stress responses. Microscopy approaches are combined with molecular, cell biology, proteomic and biochemical methods.


We are working on model plants (Arabidopsis thaliana, Nicotiana benthamiana) and the research output is translated to applications on agronomically important crops (Medicago sativa, Hordeum vulgare). Research in CRH is spearheaded by an advanced microscopy facility manpowered by experts in modern,
quantitative digital imaging.

 

  • Confocal Laser Scanning Microscopy

    Cell signaling has inherent importance to extracellular perception but also underlies cell morphogenesis and tissue development. Confocal laser scanning microscopy allows us to define subcellular localization and relationships between signaling proteins, cytoskeletal elements and endomembrane compartments. The following ZEISS product is available to work on this topic: LSM 710. The system operates with Zeiss Zen 2012 (black version) software with built in tools for quantitative FRAP, FRET and co-localization.

    • Microscopic phenotyping for plant developmental biology
    • Characterization of plant cell form and tissue architecture
    • Multicolor immunofluorescence localization of multiple antigens in various fixed plant samples (e.g. colocalization of signaling proteins with cytoskeleton)
    • Live imaging of genetically encoded fluorescent protein fusions with different spectral properties

    Arabidopsis primary root (left) and Arabidopsis lateral root primordium (right)

    Knowhow/Expertise

    • Fixed or living sample preparation and multicolor imaging
    • Quantitative colocalization between different fluorophores (Pearson’s and Manders’ coefficients)
    • Post-acquisition analysis (3-D volume rendering, fluorescence intensity profiling, orthogonal projection)
    • High resolution live cell imaging
    • 2-D time lapsed microscopy
    • Quantitative analysis of fluorescence recovery after photobleaching

    Quantitative CLSM applications in fixed and living samples

  • Spinning Disc Microscopy

    Live Cell Imaging at video rates

    Intracellular architecture may be remodeled within fractions of a second. For example membrane-derived vesicles may translocate at several µm/sec, prompting their visualization through a fast imaging platform. For such purposes we use a specially designed spinning disc platform which is able to track fast moving structures. The following ZEISS product is available to work on this topic: Cell Observer SD spinning disc system with dual camera acquisition based on the fastest EM CCD of the market, Evolve 512D from Photonics. The system is operated through Zeiss Zen 2012 (blue version) software with built in capacities for temporal and spatial quantitative analyses.

    Topic Areas

    • Quantitative tracking and analysis of fast vesicular trafficking, endocytosis and membrane recycling during diffuse or polar plant cell growth
    • Documentation and analysis of fast dynamics of actin microfilaments and microtubules during conditional, developmental or cell cycle related rearrangements in plant cells
    • Video rate imaging of conditional or developmental subcellular translocation of signaling proteins

    Knowhow/Expertise

    • Quantitative multiple tracking and characterization of different endosome types (velocity, trajectories, range of motion)
    • 2-D and 3-D imaging and analysis of fast cytoskeletal dynamics (e.g. assembly and dynamics of mitotic spindle)
    • Dynamic, simultaneous co-visualization of differentially labeled intracellular structures
    • Quantitative fluorescence recovery after photobleaching analysis of fast events (e.g. diffusion/ incorporation of fluorescent styryl dyes in the plasma membrane)
  • Wide-Field Light Microscopy

    Although CLSM is the golden standard for fluorescence imaging, conventional epifluorescence, widefield platforms are still in vigorous use. From the visualization of chromogenic cytochemical reactions to high resolution differential interference contrast imaging of living or fixed plant tissues, the CRH hosts the appropriate tool. Moreover, epifluorescence imaging can reach some CLSM standards through powerful, iterative deconvolution algorithms. The following ZEISS product is available to work on this topic: AxioImager M2 upright microscope with motorized stage and dipping objective. The microscope is operated through Zeiss Zen 2012 (blue version) with built in plugin for automatic or custom deconvolution.

    Topic Areas

    • Documentation of cell- and tissue- specific gene expression
    • Visualization of cell- and tissue- specific reactive oxygen species production
    • Routine application of conventional immunofluorescence combined with 2-D and 3-D deconvolution
    • Differential interference contrast documentation of plant cell division, growth and differentiation

    Knowhow/Expertise

    • Preparation of plant samples expressing GUS-promoter fusions for visualization of cell- and tissue- specific gene expression
    • Preparation of samples for fluorescent or chromogenic staining of reactive oxygen species
    • Time-lapsed differential interference contrast (DIC) imaging
    • Clearing and DIC documentation of plant tissue patterning
    • 2-D and 3-D image deconvolution of fluorescently (immuno) labeled samples
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