biotech quizerino for flow cytometry 2

Question 1

What are monocytes?

Answer 1

1 - Types of leukocytes that originate from the bone marrow.
2 - They constitute the basic cellular components of the peripheral blood, make up 5-10%.
3 - after a short developmental stage of about 1-3 days, they circulate briefly in the blood and migrate into tissues where they assume their ultimate role as macrophages.

Question 2

What are macrophages?

Answer 2

1 - Specific type of leukocyte that are specialized for tissue specific functions such as clearance of cellular debris, immune surveillance and the inflammatory response.
2 - Produced through differentiation from monocytes.
3 - Once differentiated, lose the ability to replicated and thus, need to be replenished.

Question 3

How can monocytes be induced to differentiate into macrophages.

Answer 3

1 - Treat with PMA
2 - Treat with Vit D3
(treat THP-1; human monocytic cell leukemia cell line)

Question 4

Describe how the phenotypic properties of monocytes and macrophages differ in terms of the analysis we will be doing.

Answer 4

Phagocytic capacity of THP-1 cells increases with differentiation due to an overall increase in cell size and volume.
1 - Macrophages have a larger observable side scatter (i.e. granularity/density) vs. monocytes and forward scatter. FL3
2 - increase in CD14 ad CD4 expression in macrophages (relative to monocytes) - FL3 channel
3 - Increased autofluorescence in macrophages relative to monocytes (since more lysosomes) - FL1
4 - Increased lysosome concentration in the macrophages due to increased cytoplasm volume - FL3