Department of Cell Biology

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.

• 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
  

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.

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.

• 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