SCYM Data Acquisition & Analysis 4

Question 1

What is the effect of overcompensation in a flow cytometry experiment?

Accepted Answer

The negative population is pulled below the baseline (false negative shift)

Answer

Overcompensation subtracts too much signal from the channel, causing the unstained population to appear with lower fluorescence than the instrument baseline.

Question 2

A log-scale biaxial plot in flow cytometry is preferred over a linear scale when displaying fluorescence data because:

Accepted Answer

It allows simultaneous visualization of dim and bright populations spanning several decades

Answer

Logarithmic display accommodates the wide dynamic range of fluorescence intensity, enabling dim and bright populations to be resolved on the same plot.

Question 3

Which analysis approach is BEST suited for identifying unexpected cell populations without prior hypothesis?

Accepted Answer

Unsupervised high-dimensional clustering algorithms (e.g., FlowSOM, t-SNE, UMAP)

Answer

Unsupervised algorithms explore all parameters simultaneously and can reveal novel or unexpected populations that manual gating strategies might miss.

Question 4

During acquisition, a sudden increase in event rate without a corresponding increase in sample volume most likely indicates:

Accepted Answer

A clog partially releasing and causing a bolus of cells

Answer

Partial clogs can trap cells and then release them suddenly, causing a transient spike in event rate that skews data.

Question 5

When designing a multicolor flow cytometry panel, bright fluorochromes should be paired with:

Accepted Answer

Lowly expressed or dim antigens to maximize sensitivity for those targets

Answer

Bright fluorochromes provide enough signal to detect dimly expressed antigens above background, while dimmer fluorochromes are sufficient for highly expressed antigens.

(SCYM) Specialist in Cytometry Practice Test

(SCYM) Specialist in Cytometry Practice Test

SCYM Data Acquisition & Analysis 4