What is the purpose of a Fluorescence Minus One (FMO) control in flow cytometry?

What is the purpose of a Fluorescence Minus One (FMO) control in flow cytometry?

• A. To measure the intensity of flurochromes in isolation
• B. To identify and gate cells by excluding one flurochrome
• C. To establish baseline fluorescence for all samples
• D. To determine the total amount of flurochrome in a panel

Answer: (B)

A Fluorescence Minus One (FMO) control is utilized in flow cytometry to set a precise gating strategy by specifically excluding one fluorescent marker from the panel. The primary purpose of this control is to determine the contribution of a single fluorochrome to the overall fluorescence signal in a multi-parameter analysis. In practice, when creating gates in flow cytometry, the FMO control allows researchers to discern whether the signal observed from a particular population of cells is genuinely positive for that specific marker or if it is simply due to background noise or spillover from other fluorescent dyes in the panel. Essentially, it helps in defining the threshold that distinguishes between positive and negative populations for the marker that has been excluded. In contrast, the other choices do not accurately capture the primary usage of the FMO control. Measuring the intensity of fluorochromes in isolation refers to a different type of control that does not relate specifically to gating. Establishing a baseline fluorescence for all samples is not the focus of an FMO control, as it is more about filtering out interference rather than creating a general baseline. Finally, determining the total amount of fluorochrome in a panel does not align with the purpose of an FMO control, which is focused on assessing the influence

A Fluorescence Minus One (FMO) control is utilized in flow cytometry to set a precise gating strategy by specifically excluding one fluorescent marker from the panel. The primary purpose of this control is to determine the contribution of a single fluorochrome to the overall fluorescence signal in a multi-parameter analysis.

In practice, when creating gates in flow cytometry, the FMO control allows researchers to discern whether the signal observed from a particular population of cells is genuinely positive for that specific marker or if it is simply due to background noise or spillover from other fluorescent dyes in the panel. Essentially, it helps in defining the threshold that distinguishes between positive and negative populations for the marker that has been excluded.

In contrast, the other choices do not accurately capture the primary usage of the FMO control. Measuring the intensity of fluorochromes in isolation refers to a different type of control that does not relate specifically to gating. Establishing a baseline fluorescence for all samples is not the focus of an FMO control, as it is more about filtering out interference rather than creating a general baseline. Finally, determining the total amount of fluorochrome in a panel does not align with the purpose of an FMO control, which is focused on assessing the influence