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Written by Tim Bushnell, PhD Flow cytometry is the science of measure the physical and biochemical processes on cells and cell-like particles. This analysis is performed in an instrument called the flow cytometer.  FACS Analysis is the short-hand expression for this type of cell analysis The term FACS stands for Fluorescent Activated Cell Sorting, a…

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Written by Tim Bushnell, PhD Every fluorophore has a unique excitation and emission profile which is usually displayed on a spectral viewer, or spectral graph. The combination of the excitation and emission profiles is the fluorophore’s spectral profile. Every fluorophore has a peak excitation wavelength (the wavelength at optimal excitation) and a peak emission wavelength…

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Written by Tim Bushnell, PhD PBS is the acronym for phosphate buffered saline. Phosphate buffer is one of the most common buffers used in biological research.  The phosphate serves as a buffer to keep the pH constant, while the saline is referencing the osmolarity.  Additional ions such as Ca2+ or Mg2+ , energy sources like…

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Written by Tim Bushnell, PhD What is autofluorescence? Autofluorescence is the term given to describe the natural fluorescence that occurs in cells. The common compounds that give rise to this fluorescence signal include cyclic ring compounds like NAD(P)H, Collagen, and Riboflavin, as well as aromatic amino acids including tyrosine, tryptophan, phenylalanine. These compounds absorb in…

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Written by Tim Bushnell, PhD Dynamic range is the total range of fluorescent values obtained from a particular flow cytometry assay. It is defined as the ratio of the largest possible fluorescent signal to the smallest possible fluorescent signal. The dynamic range can vary based on the application. For example, a cell cycle assay may…

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Written by Tim Bushnell, PhD The Fluorescence Minus One Control, or FMO control is a type of control used to properly interpret flow cytometry data.  It is used to identify and gate cells in the context of data spread due to the multiple fluorochromes in a given panel. An FMO control contains all the flurochromes…

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