Lecture 8: Flow Cytometry Flashcards
What is forward scatter?
The amount that the laser bends round the cell, measures relative size
What is side scatter?
Measures the granularity by how much light is bounced off to the side
What can be measured by flow cytometry?
Cell size, granularity, cell counting, cell markers
What is the marker for white blood cells?
CD45
What is a fluorochrome?
A molecule that can absorb light at one wavelength and emit it at another
Give two examples of fluorochromes
FITC- green, and phycoerythrin- orange
What cell populations can be detected in a lysed whole blood sample?
Lymphocytes, granulocytes, monocytes, some RBCs and platelets
Why does a whole blood sample need to be lysed?
There are many more RBCs than WBCs so they need to be lysed so they don’t interfere with the detection of WBCs
What can immunofluorescent labelling be used for?
To detect cell surface molecules or intracellular proteins, especially cytokines
Why can the % cells in a sample be abnormal when the cell count is normal?
% is relative to percentages of other cell types in the sample
What does the flow cytometry graph of someone who has chronic myeloid leukaemia look like?
Increase in number and relative % of neutrophils
What is chronic myeloid leukaemia treated with?
Tyrosine kinase inhibitors
What is compensation?
There is an overlap between wavelengths of different fluorochromes so the machine needs to be programmed to ignore these overlaps to prevent false positives
What is the marker for memory T cells and naive T cells?
Memory- CD45RO. Naive- CD45RA
