Analysis of viable and nonviable cells
AssigneeSageMedic Corporation
InventorsEkaterina Moroz, Christian Apfel, Kraig K. Anderson
Life of this patent
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Abstract
Provided are methods, for selectively analyzing a cell sample. Viable cells are dyed with a membrane-permeable fluorescent dye, and nonviable cells are dyed with a membrane-impermeable fluorescent quenching dye. The cells are illuminated to cause fluorescent emission from the membrane-permeable fluorescent dye in the viable cells and the membrane-impermeable fluorescent quenching dye in the non-viable cells. The cells are then quenched for at least a portion of fluorescence of the membrane-permeable fluorescent dye in the nonviable cells by the membrane-impermeable fluorescent quenching dye. The cells are then analyzed for viable and nonviable cells.
Claims22
A method for selectively analyzing a sample for viable cells and nonviable cells, comprising:
- providing a sample comprising one or more viable cells and/or one or more nonviable cells in a cell culture device comprising a window, the viable and/or nonviable cells capable of forming three-dimensional micro-tissue;
- culturing the one or more viable cells in the sample under cell culture conditions;
- contacting the sample with a membrane-permeable fluorescent dye and a membrane-impermeable fluorescent quenching dye, the contacting effective to cause:
- the one or more viable cells and/or the one or more nonviable cells to be dyed by the membrane-permeable fluorescent dye, and the one or more nonviable cells to be dyed with the membrane-impermeable fluorescent quenching dye, the membrane-permeable fluorescent dye characterized by a first fluorescent excitation band and a first fluorescence emission band and the membrane-impermeable fluorescent quenching dye characterized by a second fluorescent excitation band and a second fluorescence emission band;
- and
- illuminating the sample under conditions effective to cause:
- emitting of the first fluorescence emission band from the one or more viable cells dyed by the membrane-permeable fluorescent dye, emitting of the second fluorescence emission band from the one or more nonviable cells dyed by the membrane-impermeable fluorescent quenching dye, and quenching of at least a portion of fluorescence of the membrane-permeable fluorescent dye in the one or more nonviable cells by the membrane-impermeable fluorescent quenching dye;
- imaging the viable and/or nonviable cells through the window using confocal fluorescence imaging microscopy;
- and
- selectively analyzing the sample for:
- a presence of the one or more viable cells according to the first fluorescence emission band and/or a presence of the one or more nonviable cells according to the second fluorescence emission band.
Dependencies are inferred from the claim text.