Staff: Małgorzata Całka, Miguel Angel Lermo Jimenez (PhD student), Maciej Krupa (PhD student), Natalia Nowak, Błażej Ruszczycki, Hanna Sas-Nowosielska, Małgorzata Śliwińska, Artur Wolny

The Laboratory is established as a core facility providing a spectrum of microscopic techniques dedicated to functional and structural studies of biological samples. It is equipped with optical and electron microscopes enabling application of various imaging methods including time-lapse, multi-dimensional (confocal, multiphoton, deconvolution), time-resolved  (fluorescence  lifetime  and  fluorescence  correlation  spectroscopy), multispectral and high-resolution (STORM, EM, ISM) microscopy of live and fixed material. These studies are supported by image analysis and visualization algorithms, developed by the group to derive quantitative results from obtained data. Laboratory provides equipment and expertise to support a wide range of research projects concerning e.g. neuron architecture in physiological and pathological processes, cell motility, structure and dynamics of cell membranes, mitochondrial dynamics and analysis of protein dynamics and interaction in living cells.

Techniques available in the lab:

• Three dimensional correlative light and electrone microscopy (CLEM) – offers the visualization of the sample with high resolution using scanning electrone microscope toghether with multichannel imaging of fluorescently labeled intracellular structures obtained using confocal micrsocopy.
• Fast imaging of dynamic cellular processes – spinning disk confocal enables three dimensional imaging of the specimen with the speed of several frames per second. Dynamics of different organelles (e.g. mitochondria, nucleus) or molecular interactions (FRET) could be studied during live cell imaging performed with this technique.
• In vivo imaging – using the microscope with two-photon excitation of fluorescence. The studies include characterization of the neuronal network in the brain of rodents. Obtained images can be compared with the electrophysiological activity of the selected area of tissue.
• Visualization of the molecular interactions – thanks to the coupling of the confocal microscope with the time-resolved single photon detectors there is a possibility to measure  fluorescence lifetime (FLIM) as well as diffusion of particles using fluorescence correlation spectroscopy (FCS). Different oligomerization states, complex formation as well as cellular concentrations of proteins could be measured with this technique.
• Superresolution microscopy – with the set of techniques it is possible the break the diffraction limit of the conventional light microscopy. Our activities involve construction, optimization and application of techniuqes such as PALM/STORM or SIM.
• Live cell imaging using widefield microscopy – enables multichannel observations of living cells using transmitted light as well as with fluorescence method. More information about Laboratory on the web

Selected publications:

Michalska B.M., Kwapiszewska K., Szczepanowska J., Kalwarczyk T., Patalas-Krawczyk P., Szczepański K., Hołyst R., Duszyński J., Szymański J. (2018) Insight into the fission mechanism by quantitative characterization of Drp1 protein distribution in the living cel. Scientific Reports, 8, 8122 

Sas-Nowosieska H., Bernaś T. (2016) Spatial relationship between chromosomal domains in diploid and autotetraploid Arabidopsis thaliana nuclei. Nucleus, 7(2): 216-31.

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.