A student performed a flow cytometry experiment to label mouse spleen cells with fluorochrome conjugated monoclonal antibodies against mouse CD3 and CD4. The labelled cells were run through a flow cytometer and analysed. The result is presented below.

Based on this flow cytometry result, what is the most appropriate description of the cells shown in the lower left quadrant?

Question

A student performed a flow cytometry experiment to label mouse spleen cells with fluorochrome conjugated monoclonal antibodies against mouse CD3 and CD4. The labelled cells were run through a flow cytometer and analysed. The result is presented below.

Based on this flow cytometry result, what is the most appropriate description of the cells shown in the lower left quadrant?

BlackTom AI · Accepted Answer

To understand the quadrant, recall what the axes represent: CD3 on the y-axis and CD4 on the x-axis. In flow cytometry, cells that are CD3+ (T cells) appear above the horizontal line, and cells that are CD4+ appear to the right of the vertical line. Conversely, cells that are CD3- and CD4- fall in the lower left quadrant.

Option 1: "They are red blood cells". Red blood cells generally lack surface markers like CD3 and CD4 that are used to identify lymphocytes in this assay, and they typically do not cluster in this kind of antibody-stained flow plot. So this is not a correct interpretation of the quadrant shown.

Option 2: "They are double negative spleen cells". Double negative means CD3- and CD4-; these cells would appear in the lower left quadrant of a plot with CD3 vs CD4 axes. That corresponds to the described reading of the plot, making this the most accurate description.

Option 3: "They are dendritic cells". Dendritic cells can express various markers, but the standard gating here is CD3 and CD4. Since dendritic cells are not defined by CD3 and CD4 negativity here, this option is not specifically supported by the given quadrant, and it’s less precise than the double-negative designation.

Option 4: "They are B cells". B cells are typically CD3- but can be CD4- or CD4+, depending on maturation and subset, but the most specific and direct description for cells in the CD3- CD4- quadrant is double-negative cells rather than declaring them B cells.

Option 5: "They are not T cells". While CD3- CD4- cells are not T cells, this statement is too broad and nonspecific. The question asks for the most appropriate description; saying they are not T cells is less descriptive than specifically calling them double negative spleen cells.

In summary, the lower left quadrant corresponds to cells that lack both CD3 and CD4 expression, which is best described as double negative spleen cells. The other options either mischaracterize the marker expression or are too vague to be the best answer in this context.

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