Biomaterial Database

Fluorescent erythrocyte ghosts as standards for quantitative flow cytometry. 1995

S K Doberstein, and G Wiegand, and L M Machesky, and T D Pollard

Department of Cell Biology and Anatomy, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA.

We report here a quick and inexpensive method for preparing standards of known fluorochrome content for calibration and quantitation of flow cytometry fluorescence signals. Erythrocyte ghosts prepared by hypotonic lysis are filled with solutions containing fluorescently labeled dextran. Standards prepared by this technique have a narrow range of fluorescence and a linear response of fluorescence to fluorochrome content up to 2 x 10(6) fluorochrome molecules/cell. The volume of ghost standard particles is roughly 70 femtoliters (fl)/cell. The fluorescence of ghost standards is nearly identical to that of commercially available microbead standards of similar fluorochrome content. Ghost standards have stable fluorescence for at least 3 weeks at 4 degrees C. These standards can be made with any fluorochrome or combination of fluorochromes over a wide concentration range.

UI	MeSH Term	Description	Entries
D004910	Erythrocyte Membrane	The semi-permeable outer structure of a red blood cell. It is known as a red cell 'ghost' after HEMOLYSIS.	Erythrocyte Ghost,Red Cell Cytoskeleton,Red Cell Ghost,Erythrocyte Cytoskeleton,Cytoskeleton, Erythrocyte,Cytoskeleton, Red Cell,Erythrocyte Cytoskeletons,Erythrocyte Ghosts,Erythrocyte Membranes,Ghost, Erythrocyte,Ghost, Red Cell,Membrane, Erythrocyte,Red Cell Cytoskeletons,Red Cell Ghosts
D005434	Flow Cytometry	Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.	Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005453	Fluorescence	The property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D016650	Fluorescein-5-isothiocyanate	Fluorescent probe capable of being conjugated to tissue and proteins. It is used as a label in fluorescent antibody staining procedures as well as protein- and amino acid-binding techniques.	FITC,5-Isothiocyanatofluorescein,Fluorescein (5 or 6)-Isothiocyanate,Fluorescein-5-isothiocyanate Hydrochloride,5 Isothiocyanatofluorescein,Fluorescein 5 isothiocyanate,Fluorescein 5 isothiocyanate Hydrochloride,Hydrochloride, Fluorescein-5-isothiocyanate