The workshop will cover the following aspects -
- Basics of flow cytometry and its applications
- Sample preparation
- Analysis and interpretation of acquired data
The modules offered are discussed in detail below, and any one of the standard flow cytometry assays shall be demonstrated.
Hematopoietic and Mesenchymal Stem Cells
Enumeration of CD34+ hematopoietic stem and progenitor cells in bone marrow, cord blood and mobilized peripheral blood. We will discuss techniques on identifying mesenchymal stromal cells and tumor stem cells.
DNA Content and Cell Cycle Analysis
Measurement of cellular DNA content, cell cycle distribution, CDKs analysis, and effect of drugs on cell cycle will be discussed and demonstrated.
Apoptosis and Necrosis Detection
We will discuss the differences between apoptosis and necrosis, and demonstrate assays for Annexin V, caspase activation, apoptosis protein markers viz., Bax, Bcl-2, p53.
Stem Cell Assays and Phenotype Analysis
We will demonstrate and discuss sample processing and staining protocols used for phenotype analysis of hematopoietic malignancies and cell surface marker analysis of tumor stem cell antigens
Detection of ROS formation, hypoxia signaling, mitochondrial membrane potential, intracellular glutathione content, mitochondrial superoxide generation and viability