Methods in Live Cell Imaging

Subject description

Introduction of current methods, which enable observation and analysis of events appearing in live cells. The emphasis is on methods of light microscopy, which enable long-term maintaining and observation of cells in a microscope (temperature, pH and humidity controlled environment, time laps imaging, relocation of cells with collocate grids…). The mechanisms and the use of fluorescence labels (GFP, DiI, choleratoxin…), will be introduced to follow cell migration, transformation during regrowth of in vitro injuries, intercellular contact formation, rearrangement of molecules during cell differentiation, molecular dynamics in membranes, intercellular communication, cell response to drugs and toxins…) During excitation of fluorochromes the free radicals are produced, which can damage cells and cause fading of labelled structures. A survey of methods for decreased phototoxicity will be given (spinning disc microscopy, CLEM, antioxidative agents) and of possible artefacts as a result of phototoxicity.

The subject is taught in programs

Objectives and competences

  • acquaintance to methods of cell biology (time lapse imaging, fluorescence labelling of molecules in live cells)
  • ability of interpretation of processes in live cells (reorganisation of cytoskeleton in migrating cells, vesicular transport and molecular dynamics in membranes)
  • understanding of the cell dynamics in in vitro conditions (epithelio – mesenchymal transition, intercellular contact formation…)
  • understanding the reasons for the artefacts caused by fototoxisity as a result of illumination of photochromes

Teaching and learning methods

Lectures on theoretical principles, practical work in laboratory, preparation of biological specimens, microscopy, analysis of micrographs, seminar work, consultations

Expected study results

Knowledge and understanding:


The student

 – understands the dinamics of cell structures

 – is familiar with basic mechanisms of cell functioning

 – is familiar with methods of live cell labelling

 – is familiar with reasons of photodamage and resulted artefacts

Basic sources and literature

1. Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. (2008) Molecular biology of the cell, 5th edition.

   2. revijalni članki s področja, tekoča periodika, druga učna gradiva

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