
Learn the fundamentals of flow cytometry, including how a flow cytometer analyzes light from single cells and measures forward scatter, side scatter, and fluorescence for immunophenotyping and cell cycle analysis.
Cytometry measures physical and chemical properties of a single cell, and flow cytometry applies these measurements in a fluid stream using a flow cytometer.
Understand how a flow cytometer converts samples into a single cell suspension, uses hydrodynamic focusing to align cells, and measures size and fluorescence via detectors.
Principle of flow cytometry analyzes one cell at a time as it interacts with a laser, yielding forward scatter for size, side scatter for granularity, and fluorescence.
Perform a virtual flow cytometry experiment on PBMCs, labeling T cells with CD3 and B cells with CD19, then analyze FSC, SSC, and fluorescence to identify lymphocytes, monocytes, and granulocytes.
Apply flow cytometry to assess live and dead cells after anticancer drug treatment, using propidium iodide or 7-AAD staining and gating to quantify live and dead percentages.
Explore the analysis of apoptosis by annexin A5–PI flow cytometry, detailing phosphatidylserine externalization, early and late apoptotic stages, and how to interpret quadrants for live, early apoptotic, and dead cells.
Analyze the cell cycle by flow cytometry to measure DNA content with a DNA binding dye, distinguishing G1, S, and G2/M phases and calculating their cell fractions.
Learn how immunophenotyping uses flow cytometry and antibodies to define immune cell phenotypes, identifying T cells, B cells, regulatory T cells, monocytes, and NK cells in PBMCs through sequential gating.
Apply intracellular staining in flow cytometry to detect internal markers like interferon gamma and Foxp3, using fixation, permeabilization, and antibody staining to generate fluorescence.
Explore the diverse applications of flow cytometry, from cancer diagnosis and immunophenotyping to stem cell analysis, circulating tumor cells, membrane potential, ploidy, and environmental and plant studies, plus cytokine assays.
Flow cytometry is a modern and potent tool of life sciences and biological sciences. Whenever I think about this technology and its applications, I get mesmerized by the fact a single cell can be analyzed so quickly for its physical and chemical properties and this process can be repeated for millions of cells. This introductory course will help you to understand the meanings of words like cytometry immunophenotyping, hydrodynamic focusing and many more. This course is very suitable even for those students who have just begun their journey of experimental biology.
This course starts its journey with the discussions on basic principle of flowcytometry, the functioning of instrument and definitions of basic parameters like FSC, SSC and fluorescence signal. After the fundamental discussion this course takes you through various important applications of flowcytometry like live dead cell analysis, apoptosis analysis, cell cycle analysis, immunophenotyping and intracellular marker analysis etc.
The best part about this course is that to explain different applications the help of virtual experiments has been taken and original flowcytometry data and graphs have been used.
Lets Enjoy this introductory journey of flowcytometry with me and add the knowledge of this important analytical tool in your academic experience.