Methods and compositions for rapid staining of nucleic acids in whole cells
AssigneeCoulter International Corp.
InventorsChiranjit Deka, Kristie M. Gordon, Ravinder Gupta, Allan Horton
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Abstract
A rapid fluorescence staining method for facilitating flow cytometry analysis of reticulocytes is described. The method comprises contacting cells with a cocktail containing a detergent, sphering agent, and a cell impermeable dye, such as TO-PRO-3, for about one minute. Advantageously, the inventors have found that the cocktail permits the dye to penetrate the cell membrane rapidly.
Claims16
A method for facilitating rapid transport of dye molecules through a cell membrane comprising the steps of contacting a cell sample with a mixture consisting of a cell membrane impermeant dye, a non-ionic detergent and a sphering agent.
A method for staining nucleic acids in a cell using a cell membrane impermeant dye, said method comprising the steps of:
- (a) providing a sample comprising a cell;
- and
- (b) contacting the sample with a mixture consisting of a non-ionic detergent, a sphering agent and a cell membrane impermeant dye for nucleic acids.
A method for rapid analysis of reticulocytes with a cell impermeable dye comprising the steps of:
- (a) contacting a cell sample containing reticulocytes with a mixture consisting of a cell impermeable dye, a non-ionic detergent and a sphering agent;
- and (b) analyzing the cells by automated flow cytometry to detect the presence of stained reticulocytes.
A composition which facilitates transport of cell impermeable dye molecules through cell membrane;
- said composition consisting of a non-ionic detergent, a sphering agent, and cell impermeable dye molecule.
Dependencies are inferred from the claim text.