18 Questions
What is the purpose of the forward scatter detectors in a flow cytometer?
To measure the size of the cell, with larger cells scattering more light.
How does the complexity of the cell's internal features affect the side scatter intensity?
The greater the complexity, the greater the deflection of laser light into the side aperture, resulting in higher side scatter intensity.
What role do optical filters and dichroic mirrors play in the flow cytometry technique?
They filter and move light to the detectors.
What is the primary function of a Fluorescence Activated Cell Sorter (FACS)?
To sort cells based on their fluorescent characteristics.
What is the purpose of staining cells with fluorescence-labelled antibodies in flow cytometry?
To allow for the measurement of specific cell surface markers or intracellular proteins.
What is the outcome of the flow cytometry technique?
The measurement of the properties of each cell, including size, granularity, and fluorescent characteristics.
What is the purpose of the electrical charging in the flow cytometry technique?
To charge the droplets so that they can be directed into different containers based on their charge.
Why are red cells lysed before putting the sample in the flow cytometer?
To remove them from the analysis, allowing for the focus to be on white cells.
What is the purpose of lymphocyte gating in flow cytometry?
To select or 'gate' lymphocytes from the analysis of all white cells.
What is the purpose of using CD3 and CD19 antibodies in the staining process?
To distinguish between different types of lymphocytes, such as T cells, B cells, and NK cells.
What is the principle of the fluorescence activated cell sorter (FACS) in flow cytometry?
To separate cells based on their fluorescence properties, which are determined by the antibodies used to stain the cells.
What is the advantage of using flow cytometry in immunophenotyping?
It allows for the rapid analysis of multiple cell populations and their characteristics, enabling the identification of specific cell types and their proportions.
What are two of the most important tests that the hospital doctors perform on Patient 2?
Serum immunoglobulins and lymphocyte immunophenotyping
Why do infants typically have lower levels of IgG compared to adults?
Ig is slow to develop in children, and levels are lower as they produce their own IgG over time.
Why are monoclonal antibodies pre-labelled with different fluorescent dyes in lymphocyte immunophenotyping?
To detect different lymphocyte subsets based on their surface markers.
How can CD4 T, CD8 T, and B cells be distinguished in lymphocyte immunophenotyping?
By using monoclonal antibodies pre-labelled with fluorescent dyes that target specific surface markers.
What technique is used to detect the subsets of lymphocytes based on different surface markers?
Fluorescence-activated cell sorter (flow cytometry technique)
Why are optical filters and dichroic mirrors important in flow cytometry?
They help in separating the emitted fluorescence signals and directing them to the appropriate detectors.
Learn about the case of a 6-month-old boy with repeated infections, severe pneumonia, and middle ear infections leading to suspicion of immunodeficiency. Explore the importance of serum immunoglobulins and lymphocyte immunophenotyping tests in diagnosing immunodeficiency.
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