Automated method for cell volume determination
AssigneeOrtho Diagnostics, Inc.
InventorsW. Peter Hansen, Robert A. Hoffman, Peter J. Natale
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Abstract
Fluorescence volume exclusion signals are utilized to evaluate cell volume. Cells are suspended in a medium which is furnished with a fluorescent dye which neither penetrates nor adheres to the cells. Cells are analyzed by an optical flow cytometer, with illumination which causes the dye medium to fluoresce. Passage of a cell through the sensing zone therefore reduces the amount of fluorescent medium being illuminated, and proportionally reduces the fluorescence output signal. Fluorescence volume exclusion pulse height and area are related to cell volume.
Claims6
A method of evaluating cell volume in a sample of cells comprising the steps of:
- (a) suspending the cells of said sample in a medium including a dye which exhibits fluorescence response to select stimulation, and which is non-penetrating and substantially non-adherent to said cells;
- (b) passing said medium through an optical flow cytometry sensing zone illuminated by said select stimulation;
- (c) detecting reductions of fluorescent emissions from said sensing zone occasioned by passage of cells through said zone;
- and
- (d) evaluating cell volume based on the amplitude of said detected reductions of fluorescent emissions.
Dependencies are inferred from the claim text.