LABORATORY OF MICROSCOPIC TECHNIQUES

About the lab

Research area of ​​the laboratory includes investigation of pathomechanisms occurring at early stages of neurodegenerative disorders. Current projects are aimed at development of diagnostic options of neurodegeneration relevant for Parkinson´s disease. We also deal with possible intervention strategies to reduce the impact of a disease, or with description of mechanisms of neurodegeneration caused by ischemic damage to the central nervous system.Experimental research includes cellular modelling of neurodegenerative processes, but also animal models are used. In addition to experimental models, research involving human specimens is planning to be performed in cooperation with theoretical and clinical departments of JFMED CU.

In the field of cell research, the laboratory is focusing on the observation of changes taking place in living, both primary and commercial, cell lines. Using in vivo fluorescence microscopy, we observe morphological changes of intracellular structures. In addition, it also deals with functional changes occurring in the mitochondria and endoplasmic reticulum. Fluorescence microscopy is thus performed for the purpose of observing the dynamic rearrangement of membrane structures. It is also possible to observe rapid changes in the intracellular concentrations of signal ions, such as Ca2+.

Animal modeling of neurodegeneration as well as processing of human samples lead to the analytical part of an experiment predominantly focusing on evaluation of complex morphological changes occurring in fixed post mortem/ex vivo tissue samples. Thanks to the tunable infrared laser, we are able to visualize samples several millimeters thick, depending on the previous preparation and type of sample. Imaging of entire parts of organs, such as gastrointestinal tract, is highly relevant for creating a comprehensive insight about changes occurring in the tissue. In addition to the classic imaging using the concentration of the visualized molecule, the lab is equipped with such instruments which allow to observe changes in fluorescence lifetime. This microscopic modality is very effective mainly due to its sensitivity which can be used to reveal possible molecular interactions between examined factors.

The vision of the laboratory is to create and constantly improve the possibilities for detailed and sensitive description of changes taking place in biological systems at very early stages of development of degenerative changes leading to irreversible damage to systemic part s of the body.

Staff

  •  Head:
    RNDr. Michal Pokusa, PhD.
    michal.pokusa@uniba.sk
    +421 2633 643

  • Researcher:
    RNDr. Andrea Evinová, PhD.

Video tour

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Laboratory infrastructure

Laboratory of Microscopic Techniques with an intensive support of the Department of Pathological Physiology is equipped with the following florescence microscopes:

  1. Zeiss LSM 880 with multiphoton laser
  2. Zeiss Axio imager M.2 with apotome module
  3. Zeiss Axio observer D.1 with inverted lens and DG4 fluorescent source
  4. Zeiss Axio examiner A.1 equipped with a patch clamp micromanipulator
  5. Olympus BX51

 

In addition to microscopy, common workplaces of the institute and biomedical center offer possibilities for preparation and maintenance of cell lines. Processing of fixed histological and cell preparations for the purpose of immunodetection can be performed in a supporting biochemical laboratory.

Virtual tour

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Projects

VEGA- 1/0299/20 Study of the influence of preischemia and global ischemia of the brain on proteins of mitochondrial dynamics

VEGA- 1/0371/21 New approaches to the detection of pathological forms of alpha-synuclein in gastrointestinal tissues.

The most important publications

Pokusa M, Hajdúchová D, Menichová V, Evinová A, Hatoková Z, Kráľová-Trančíková A. Vulnerability of subcellular structures to pathogenesis induced by rotenone in SH-SY5Y cells. Physiol Res. 2021 Mar 17;70(1):89-99. Epub 2021 Jan 14. PMID: 33453717.

Pokusa M, Kráľová Trančíková A. FLIM analysis of intracellular markers associated with the development of Parkinson’s disease in cellular model. Physiol Res. 2018 Dec 31;67(Suppl 4):S673-S683. doi: 10.33549/physiolres.934054. PMID: 30607974.

Pokusa M, Kráľová Trančíková A. The Central Role of Biometals Maintains Oxidative Balance in the Context of Metabolic and Neurodegenerative Disorders. Oxid Med Cell Longev. 2017;2017:8210734. doi: 10.1155/2017/8210734. Epub 2017 Jul 2. PMID: 28751933; PMCID: PMC5511683.

Pokusa M, Hajduchova D, Buday T, Kralova Trancikova A. Respiratory Function and Dysfunction in Parkinson-Type Neurodegeneration. Physiol Res. 2020 Mar 27;69(Suppl 1):S69-S79. doi: 10.33549/physiolres.934405. PMID: 32228013.