Get University Of Missouri LIDR Immunology Core Flow Cytometry Information Sheet 2018-2024

Your sample of blood, bone marrow or tissue cells is placed in a suspension and injected into the flow cytometer machine. The cells are arranged in a single file line, and then passed in front of a laser beam, scattered light and fluorescent light. Next, the cells are counted and categorized.

There are a wide variety of applications for flow cytometry in immunology, including tracking expansion of antigen-specific T cells by running a large total number of cells to detect a small percentage of specific cells. Increased information comes from staining for multiple antigenic markers.

Traditional flow cytometers consist of three systems: fluidics, optics and electronics. The fluidics system consists of sheath fluid (usually a buffered saline solution) that is pressurized to deliver and focus the sample to the laser intercept or interrogation point where the sample is analyzed.

Flow cytometry directly detects antigens using fluorescent conjugated antibodies, each corresponding to a different protein in or on the cell surface. A key advantage of flow cytometry is that it allows researchers to simultaneously analyze many parameters in a cell population and many different cell markers.

Flow cytometry data analysis is built upon the principle of gating. Gates and regions are placed around populations of cells with common characteristics, usually forward scatter, side scatter and marker expression, to investigate and to quantify these populations of interest.

FACS: Sorting Cells based on Flow Cytometry Data FACS is a derivative of flow cytometry that adds an exceptional degree of functionality. Using FACS a researcher can physically sort a heterogeneous mixture of cells into different populations.

Flow cytometry is a powerful technique that is widely used to identify and characterize different immune cell types in heterogeneous samples. It primarily relies on the use of fluorochrome-conjugated antibodies to detect the expression of specific cell surface or intracellular antigens on single cells in suspension.