What controls are used in flow cytometry?
Controls in Flow Cytometry
- Unstained peripheral blood.
- Staining of THP-1 cells with CD11a in the presence or absence of Fc block.
- Using a viability dye to exclude dead cells.
- Secondary antibody alone control staining of porcine lymphocytes.
- Use of a biological control.
- Use of stimulated and unstimulated controls.
What is immunophenotyping in flow cytometry?
Immunophenotyping by flow cytometry is a laboratory method that detects the presence or absence of white blood cell (WBC) markers called antigens. Immunophenotyping by flow cytometry is a laboratory method that detects the presence or absence of white blood cell (WBC) markers called antigens.
Why should isotype controls be used in immunophenotyping?
Isotype controls are antibodies raised against an antigen not found on the cell type or sample analyzed. They have been developed for surface staining and their role is to ensure the observed staining is due to specific antibody binding to the target rather than an artifact.
What is a positive control in flow cytometry?
A positive control is an aliquot of the cells being studied stained with a single fluorochrome. However, be aware that it is not uncommon to include an experimental control in flow cytometry experiments in order to verify that everything is working as expected. A typical example would be a cell cycle control.
What is FMO control in flow cytometry?
Fluorescence Minus One (FMO) controls are samples stained with all the fluorophores in your panel, minus one of them. They are used to set the upper boundary for background signal on the omitted label, and thus to identify and gate positive populations in multicolor experiments.
Does flow cytometry always use antibodies?
Flow cytometry is an invaluable tool used to analyze the chemical and physical properties of cells. This laboratory technique uses an antibody conjugated with a fluorochrome for cell analysis. The most important clinical application of flow cytometry is in hematologic malignancy diagnosis.
What is a gate in flow cytometry?
Data analysis in flow cytometry 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.