Laboratory of Imaging Tissue Structure and Function
Head: Tytus Bernaś
Staff: Wojciech Brutkowski, Wanda Kłopocka, Jarosław Korczyński, Błażej Ruszczycki, Artur Wolny
The laboratory is dedicated to study of architecture and dynamics of biological structure at several levels of organization, ranging from imaging of subcellular structures to animal organs in vivo. To this end, a range of optical and electron imaging modalities are provided to collaborating scientists. The available techniques include:
• correlative microscopy (CLEM) in 3D, combining high-resolution imaging with scanning electron microscope (SEM) and physical tomography with multiplexed imaging of molecularly specific probes with optical (confocal fluorescence) tomography at low resolution. Current applications include imaging of architecture of synapses and spines in neurons.
• rapid imaging of cellular dynamics, with spinning disc confocal microscope providing 3D imaging up to 20 frames (datasets) per second. The applications include probing of cellular signaling cascades with specific FRET probes and investigation of changes of organelle (ex. nuclei and mitochondria) architecture.
• in vivo imaging, implemented with two-photon microscope. The research involves characterization of neuronal morphology in living rodent brain and its change in physiological processes. Furthermore, the equipment is used to correlate morphology of tissue with its electro- physiological activity.
• mapping of molecular interactions, performed with photon counting confocal microscope providing characterization of fluorescence life time (FLIM), polarization anisotropy and correlation (FCS). Planned applications involve probing interactions of biomolecules in cells and comprise measurements of metalloproteinase activity in situ and formation of histone-DNA complexes.
• super-resolution microscopy, comprising a set of optical imaging modalities providing higher spatial resolution than traditional, diffraction-limited microscopy. Current activities include construction, optimization and biologi- cal implementation of the techniques, whose examples are PALM/STORM and SIM.
• wide-field system for live imaging of motile cells, multifield time laps analysis in transmitted light as well as broad spectrum of fluorescence methods. Fast filter wheel sys- tem is especially suited for rapid calcium measurements. System is also equipped with optics for IRM cell adhesion imaging. More information about Laboratory on the web
Kilańczyk E., Graczyk A., Ostrowska H., Kasacka I., Leśniak W., Filipek A. (2012) S100A6 is transcriptionally regulated by β-catenin and interacts with a novel target, lamin A/C, in colorectal cancer cells. Cell Calcium, 51(6): 470-477.
Knapska E., Macias M., Mikosz M., Nowak A., Owczarek D., Wawrzyniak M., Pieprzyk M., Cymerman I.A., Werka T., Sheng M., Maren S., Jaworski J., Kaczmarek L. (2012) Functional anatomy of neural circuits regulating fear and extinction. Proceedings of the National Academy of Sciences USA, 42: 17093-17098.
Dziembowska M., Milek J., Janusz A., Rejmak E., Romanowska E., Gorkiewicz T., Tiron A., Bramham C., Kaczmarek L. (2012) Activity-dependent local translation of matrix metalloproteinase-9. Journal of Neuroscience, 32: 14538–14547.
Biegańska K., Figiel I., Gierej D., Kaczmarek L., Klejman L. (2012) Silencing of ICERs (Inducible cAMP Early Repressors) results in partial protection of neurons from Programmed Cell Death. Neurobiology of Disease, 45: 701–710.
Samluk L., Czeredyś M., Skowronek K., Nałęcz K.A. (2012) protein kinase C regulates amino acid transporter ATB, Biochem- ical and Biophysical Research Communications, 422, 64-69