Methods for identifying stem cells by detecting fluorescence of cells and syncytia
AssigneeMassachusetts Institute of Technology
InventorsElena V. Gostjeva, William G. Thilly
Life of this patent
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Abstract
Methods are disclosed for the characterization of the stage of development or pathology of a tissue sample, and for identifying pluripotent metakaryotic stem cells, comprising detecting fluorescence of cells and syncytia in fixed samples treated with a non-fluorescent Schiff's base reagent in the absence of extraneous or exogenously added fluorescent dyes.
Claims9
A method for identifying a metakaryotic stem cell of interest having a bell-shaped nucleus, the method comprising:
- a) fixing a sample of cells with an acid-alcohol mixture;
- b) treating said sample with a Feulgen's reagent with fuchsin as the Schiff's base;
- c) maintaining said sample in contact with the Feulgen's reagent in order to stain the sample in the absence of extraneous or exogenously added fluorescent dyes;
- d) exposing said stained sample to an excitation light from a light source;
- and
- e) visualizing green-light emitting fluorescent balloon-like cytoplasmic structures of a metakaryotic stem cell by fluorescence microscopy, thereby identifying the metakaryotic stem cell of interest in the sample.
A method for diagnosing preneoplasia or neoplasia in an adult mammal suspected of having preneoplasia or neoplasia, the method comprising:
- a) fixing a sample of cells obtained from the adult mammal with an acid-alcohol mixture;
- b) treating said sample with a Feulgen's reagent with fuchsin as the Schiff's base;
- c) maintaining said sample in contact with the Feulgen's reagent in order to stain the sample in the absence of extraneous or exogenously added fluorescent dyes;
- d) exposing said stained sample to an excitation light from a light source;
- and
- e) detecting green-light emitting fluorescent balloon-like cytoplasmic structures of a metakaryotic stem cell in the sample by fluorescence microscopy, whereby detecting the presence of one or more green-light emitting fluorescent metakaryotic stem cells having a bell-shaped nucleus in the absence of extraneous or exogenously added fluorescent dyes is indicative of preneoplasia or neoplasia in the adult mammal.
Dependencies are inferred from the claim text.