Zakład Biologii Komórki

Teachers:

• dr hab. Jolanta Sroka (co-ordinator)
• prof. dr hab. Jarosław Czyż, dr hab. Ewa Zuba-Surma, prof. UJ, dr Monika Rak

Participation in the practical course: 30 hr

ECTS: 3

Semester: summer (12th April - 24th May 2016)

Place: seminar class C110

Tuesday 13.30-18.00

Credit: Written test will be held to test the knowledge acquired during the course.

The course includes:

1. Tutorial on the principles of fluorescence microscopy, digital imaging and the practical use of automatic microscopes, in particular Leica DM IRE2 fluorescence microscope equipped with Leica DC350 FX cooled-CCD camera (Fluorescence microscopy). Image capture technology based on digital camera systems with charge-coupled devices (CCD) in modern optical microscopy. Collecting high quality (low noise/signal ratio) digital images. Practical use of a software-controlled CCD camera that allows collecting, organizing, and processing of digital images acquired with Leica DM IRE2 automatic fluorescence microscope equipped with Leica DC350 FX cooled-CCD camera)
2. Principles and practical use of fluorescence microscopy for cytoskeleton staining (actin and tubulin) in fixed cells. Visualisation of the differences in cytoskeleton organization between the normal and cancer cells. Nocodazole, cytochalasins and phalloidin as the agents affecting cytoskeleton organization and their use in molecular biology (staining the actin filaments with fluorochrome-conjugated phalloidin. Immunofluorescence.
3. Practical tutorial on the computer-aided analysis of time lapse video recordings acquired with automatic microscope to examine some biological processes, such as cell migration.
4. Intercellular exchange of regulatory signals and principles of its regulation. Functions of connexins and gap junctions: their role in homeostasis and pathology with special emphasis to cancer. Application of fluorescence microscopy in the visualization of intercellular exchange of small metabolites via gap junctions.
5. Harvesting of bone marrow (BM) tissue form murine bone cavities and its futher processing for fluorescent immunolabelling. BM- derived cells will be stained against specific antigens characterizing selected stem cell (SC) populations that will be eventually identified by multicolor flow cytometry.

Schedule

1. Principles and practical use of fluorescence procedures for actin and tubulin cytoskeleton staining in fixed cells

2. Principles of fluorescence microscopy, digital imaging and automatic microscopes and their practical use

3. Harvesting of bone marrow tissue from murine bone and its further processing for fluorescent labelling

4. Practical use of methods based on the computer-aided analysis of time-lapse video recording to examine biological processes (cell migration) 

5. Intercellular exchange of regulatory signals and principles of its regulation. Functions ofconnexins and gap junctions: their role in homeostasis and pathology with special emphasis to cancer