Flow Cytometry Flashcards
Define flow cytometry.
A system in which single cells (or beads) in a fluid suspension are analyzed in terms of their intrinsic light-scattering characteristics.
In flow cytometry, what helps determine what proteins are present on the cell?
Color detectors.
What parts of the cell is flow cytometry able to measure?
Cell size, granularity, surface receptors, total DNA gene expression, and intracellular proteins.
What specimens are acceptable in flow cytometry?
Whole blood - EDTA or heparin (heparin increases stability time);
Bone marrow - heparin;
Tissue samples.
Define fluorophores.
Fluorescent markers that detect expression of cellular molecules (e.g., proteins, nucleic acids) which accept light energy (laser).
What types of scatter does light at two specific angles are measured by the flow cytometer?
Forward and side scatter (intrinsic).
What extrinsic parameter does the flow cytometer used for cell detection?
Fluorescent probes.
Define gates.
A system in which single cells (or beads) in a fluid suspension are analyzed in terms of their intrinsic light-scattering characteristics.
Describe a single-parameter histogram plot.
Plots a chosen parameter (generally fluorescence) on the x-axis versus the number of events on the y-axis; thus, only a single parameter is analyzed using this type of graph.
Describe a dual-parameter dot plot.
A bivariate histogram where each dot represents an individual cell or event. Two parameters, one on each axis, are plotted against each other.
When analyzing a population of cells using a dual-parameter dot plot, the operator chooses which parameters to analyze on both the x- and y-axes, divides the dot plot into ___ quadrants, and separates the positive events from the negative events in each axis
Four.
In quadrant analysis, the first quadrant consists of what types of cells?
Positive for fluorescence on the y-axis and negative for fluorescence on the x-axis.
In quadrant analysis, the second quadrant consists of what types of cells?
Positive for fluorescence on both the x- and y-axes.
In quadrant analysis, the third quadrant consists of what types of cells?
Negative for fluorescence on both the x- and y-axes.
In quadrant analysis, the fourth quadrant consists of what types of cells?
Positive for fluorescence on the x-axis and negative for fluorescence on the y-axis.
