Lymphocyte Characterisation Flashcards
What is the function of PMA/Ionomycin?
Induces direct signal transduction
What is the function of Poly I:C?
A dsRNA synthetics analogue
What are the most prevalent cells in the blood and spleen?
Red blood cells
What makes red blood cells easy to eliminate from the cell preparations?
They are more sensitive to osmotic lysis than other cells
Why do we eliminate the red blood cells from the sample?
To see clearly the other cell types
What happens when a mixed population of cells are exposed to either distilled water or dilute salt solution for a very short time?
The red blood cells will burst but other cells will remain intact
What are some applications of flow cytometry?
Detect different cell types based on:
1. Physical properties
2. Based on cell surface markers expressed
3. Functional readouts (cytokine production)
How does flow cytometry work?
A mixed cell population is passed one cell at a time past both light and laser beams. The light beam is scattered by the cells and different cell types have different scatter characteristics
In flow cytometry, what will the amount of light getting straight through depend on?
The size of the cells (forward scatter)
In flow cytometry what will the amount of light diffracted off at angles depend on
The internal granularity of the cells (side scatter)
What colour will the supernatant be if red blood cell lysis was efficient? and why?
Red
Due to haemoglobin release
What is Brefeldin A?
Blocks protein export from the golgi resultin in intracellular accumulation of cytokines
What can FACS measure?
A percentage rather than the actual number of cells
What does fix A do?
Kills cells
What does perm B do
Makes holes in membranes of cells
