Carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) is an

amine In chemistry, amines (, ) are compounds and functional groups that contain a basic nitrogen atom with a lone pair. Amines are formally derivatives of ammonia (), wherein one or more hydrogen atoms have been replaced by a substituen ...

-reactive,

cell Cell most often refers to: * Cell (biology), the functional basic unit of life Cell may also refer to: Locations * Monastic cell, a small room, hut, or cave in which a religious recluse lives, alternatively the small precursor of a monastery w ...

-permeable

dye A dye is a colored substance that chemically bonds to the substrate to which it is being applied. This distinguishes dyes from pigments which do not chemically bind to the material they color. Dye is generally applied in an aqueous solution an ...

generally used in animal cell proliferation research. CFDA-SE is a modified CFSE with two

hydroxyl In chemistry, a hydroxy or hydroxyl group is a functional group with the chemical formula and composed of one oxygen atom covalently bonded to one hydrogen atom. In organic chemistry, alcohols and carboxylic acids contain one or more hydroxy ...

groups on its fluorescein moiety replaced with

acetates An acetate is a salt formed by the combination of acetic acid with a base (e.g. alkaline, earthy, metallic, nonmetallic or radical base). "Acetate" also describes the conjugate base or ion (specifically, the negatively charged ion called an a ...

. This change renders the molecule more

hydrophobic In chemistry, hydrophobicity is the physical property of a molecule that is seemingly repelled from a mass of water (known as a hydrophobe). In contrast, hydrophiles are attracted to water. Hydrophobic molecules tend to be nonpolar and, th ...

and cell-permeable at the expense of its fluorescence property. After entering cells by diffusion, CFDA-SE is cleaved by

intracellular This glossary of biology terms is a list of definitions of fundamental terms and concepts used in biology, the study of life and of living organisms. It is intended as introductory material for novices; for more specific and technical definitions ...

esterase An esterase is a hydrolase enzyme that splits esters into an acid and an alcohol in a chemical reaction with water called hydrolysis. A wide range of different esterases exist that differ in their substrate specificity, their protein structure, ...

enzymes to form CFSE. As its reactive succinimidyl ester group is unmodified, CFDA-SE also

covalently A covalent bond is a chemical bond that involves the sharing of electrons to form electron pairs between atoms. These electron pairs are known as shared pairs or bonding pairs. The stable balance of attractive and repulsive forces between atoms ...

binds to

lysine Lysine (symbol Lys or K) is an α-amino acid that is a precursor to many proteins. It contains an α-amino group (which is in the protonated form under biological conditions), an α-carboxylic acid group (which is in the deprotonated −C ...

residues and other amine sources like CFSE. If a stained cell divides, the dye is divided equally between the two daughter cells, resulting in both new cells having a CFDA-SE

concentration In chemistry, concentration is the abundance of a constituent divided by the total volume of a mixture. Several types of mathematical description can be distinguished: '' mass concentration'', ''molar concentration'', ''number concentration'', an ...

approximately 50% that of the mother cell. A cell stained with CFDA-SE can be kept in

culture Culture () is an umbrella term which encompasses the social behavior, institutions, and norms found in human societies, as well as the knowledge, beliefs, arts, laws, customs, capabilities, and habits of the individuals in these groups.Tyl ...

for several days and fluorescence is detectable in cells following up to 8 successive cell divisions. Fluorescence is typically detected using a

flow cytometer Flow cytometry (FC) is a technique used to detect and measure physical and chemical characteristics of a population of Cell (biology), cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and inject ...

on the FL1 detector, with each resulting fluorescent peak representing another round of cell division. The area of each peak is representative of the number of cells in a given division cycle. The staining works best with relatively homogeneous cell populations. High concentrations of the dye are toxic to animal cells; however, concentrations in the region of 10

micromolar Molar concentration (also called molarity, amount concentration or substance concentration) is a measure of the concentration of a chemical species, in particular of a solute in a solution, in terms of amount of substance per unit volume of sol ...

are typically sufficient to give strong staining with minimal cell death. Most cell types will excrete a proportion of the dye over the course of the first 24–48 hours following staining. Dye levels should thereafter remain relatively stable in non-dividing cells. CFDA-SE is often erroneously referred to as CFSE, even in scientific literature.Cell Lab Quanta SC
{{webarchive , url=https://web.archive.org/web/20070315183008/http://www.beckmancoulter.com/products/instrument/cellanalysis/celllab_quanta_sc_dcr.asp , date=March 15, 2007

References

Fluorone dyes Vital stains Acetate esters Succinimides

See also

References