Institute of Molecular Genetics ASCR, v.v.i.

Biology of the Cell Nucleus

Address

Vídeňská 1083
14200 Prague
Czech Republic

Phone: +420241062219
http://www.img.cas.cz/
http://www.img.cas.cz/research/pavel-hozak/

Contacts

Head

Pavel Hozak
Phone: +420241062219
Email: hozak@img.cas.cz

Involved in

  • WG1 - Biophysics of cell and tissue structure
  • WG2 - Structural analysis of biomolecules involved in mechanobiology
  • WG3 - New methodologies to study mechanobiology of cells and tissues
  • WG4 - Mechanobiological principles of rare and common diseases

General description research focus

In diploid mammalian cells, some 6×109 base pairs of DNA fold as a nucleoprotein complex (i.e. chromatin) into higher-order arrays so as to fit in a nucleus measuring only 10 µm. The nucleus also contains machineries for transcription of genes and processing of RNA products, and for precise DNA replication, repair and recombination. Nuclear interior is therefore functionally highly compartmentalized, and the recent evidence points strongly to structure-related regulation of nuclear functions – however, the mechanisms forming the 3D-structure of the nucleus are still mostly obscure. We therefore employ a multi-disciplinary approach in order to study nuclear functions in relation to the higher-order nuclear structures, e.g. nuclear bodies, the nucleolus, and the nucleoskeleton. 

Imaging technologies available

  • Laser Scanning Confocal Microscopy (LSM / CLSM)
  • Deconvolution Widefield Microscopy
  • Total Internal Reflection Fluorescence Microscopy (TIRF)
  • Stimulated Emission Depletion Microscopy (STED)
  • Stochastic Optical Reconstruction Microscopy (STORM)
  • Fluorescence Resonance Energy Transfer (FRET)
  • Fluorescence Recovery After Photobleaching (FRAP)
  • Electron Microscopy
  • Structured llumination Imaging (SIM) technology
  • Electron tomography
  • Live cell imaging

Equipment available

http://www.img.cas.cz/core-facilities/microscopy-centre/equipment/

Expertise support provided to users

The facility provides methodological and instrumentation background for flow cytometry and fluorescence microscopy techniques. The facility is equipped with two flow cytometers with high-through automated samplers and one polychromatic high-speed cell sorter. Laboratory facilities include also AutoMACS Pro magnetic separator for automatic rapid sorting of cells. Light microscopy instrumentation includes twelve microscopy systems which range from routine fluorescence microscopes, confocal microscopes up to high-end super-resolution microscopy systems STED and N-SIM. Most of the instrumentation in the laboratory is available on the self-service basis, for trained users. Big emphasis is given not only for data acquisition, but for image reconstruction, processing and analysis as well. Laboratory is running cell sorting service. Several offline analysis workstations are also available in the facility, for analysis of flow cytometric (FlowJo) and image data (SoftWorx Suite, Imaris, LAS AF, Huygens, ImageJ, NIS Elements). 

The core facility deals with various biological samples: human and animal cell cultures, plant and animal tissues, worms, microorganisms, lipid micelles. The sample preparation techniques include routine chemical fixation and resin embedding, cryofixation using high-pressure freezing technique, freeze-substitution, plunge-freezing, cryosectioning, and immunolabeling, including simultaneous detection of multiple targets by our self-developed methods.

Research topics directly investigated in research group

(1) the relationship between nuclear compartmentalization and regulation of gene expression, (2) structure, dynamics, and function of the nucleoskeleton, which contributes to the nuclear compartmentalization, (3) functions of nuclear myosins and actins in transcription and gene expression, (4) functions of nuclear lipids, (5) development of new microscopy methods.

Access rules research groups

Open user access available under the Czech-BioImaging: https://www.czech-bioimaging.cz/czech-bioimaging-access-policy