The Course will be held on October 25 and 26^th 2012 in

the Lecture Hall of the Institute of Biology and Immunology of Reproduction

Bulgarian Academy of Sciences

73 Tzarigradsko shosse, Sofia

25.10. 2012

09.00 – 10.30 Theoretical part: Introduction to Flow Cytometry.

Demonstration of some Flow Cytometry applications for cell analysis.

Assoc. Prof. Tsvetelina Oreshkova (IBIR)

• FACSCalibur instrument overview and functioning. Measurement of physical and chemical characteristics of single cells. Specialized programs for analysis.
• Major parameters for cell characterization and analysis. Multiparametric and statistical analyses. Flow cytometry applications.
• Theoretical introduction to experiments for demonstration.

10.30 – 12.30 Practical part: – Assoc. Prof. Tsvetelina Oreshkova (IBIR)

• Sample preparation. Cells staining with dyes and fluorochrome-labeled antibodies for analyses.

Lunch

14.00 – 16.00 Flow Cytometry acquisition and analyses

• Calibration of FACSCalibur.
• Immune-phenotypisation.
• Measurement of lymphocyte proliferation.
• Measurement of apoptosis in tumor cells.

• 26.10. 2012 г.

10.30 – 12.00 Theoretical part: General approaches in the study of male gametes quality using flow cytometry.

Assist. Prof. Soren Hayrabedyan (IBIR)

• Theoretical and methodological background of research male gametes quality assessment in humans and animals.
• Study on parameters and algorithms: cell vitality, apoptotic changes, oxidative stress evaluation using generated reactive oxygen species, acrosomal status, capacitation status, mitochondrial potential, chromatin status, size and quantity of sperm cells.

Lunch

14.00 – 16.00 Practical part: Practical demonstration of flow cytometric analysis of some of the parameters listed above in animal ejaculates.

Assessment and comparison with experimentally controlled control group of cells obtained from a standard cell line.

Assist. Prof. Soren Hayrabedyan (IBIR)

• Analysis of the experimental protocols.

Practical session.