You Are Going to Use Flow Cytometry to Determine the Proportion

You are going to use flow cytometry to determine the proportion of developing B cells in the bone marrow that are immature, anergic, or mature. You have three monoclonal antibodies specific for three different B-cell surface proteins. The first has specificity for the cell-surface protein CD19, which is expressed by all developing and mature B cells; the second is specific for the Fc region of IgD; and the third is specific for the Fc region of IgM. The antibodies are conjugated to three different fluorescent tags that can be detected and distinguished by the flow cytometer.
A. Use histograms to show your analysis of CD19-positive cells and indicate which part of your histogram you would gate to analyze IgM and IgD expression. Indicate the gated population with an arrow.
B. Using a two-dimensional dot plot, compare the expression of IgD and IgM of these gated cells, and say which of these populations represents (i) immature B cells, (ii) mature B cells, and (iii) anergic B cells.

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