Method for conducting multiparametric fluorescence in situ hybridization

Claim 1

A method of simultaneously identifying and distinguishing the individual autosomal and sex chromosomes of a human karyotype which comprises the steps: (a) contacting a preparation of said chromosomes, in single-stranded form, under conditions sufficient to permit nucleic acid hybridization to occur with a set of combinatorially labeled oligonucleotide probes, each member thereof: (i) having a predetermined fluorescent label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal and sex chromosome of said human karyotype; wherein said contacting thereby results in each autosomal or sex chromosome of said preparation being hybridized to at least one member of said set of probes; (b) for each chromosome of said preparation hybridized to at least one member of said set of probes, employing an interferometer to detect and identify the predetermined label of the hybridized probe member(s) to correlate the identity of the label with the identity of the autosomal and sex chromosomes of said human karyotype, thereby identifying the chromosome hybridized to said probe member(s); and (c) repeating step (b) until each autosomal and sex chromosome of said human karyotype has been identified in said preparation.

Claim 2

A method of simultaneously identifying and distinguishing the individual autosomal and sex chromosomes of a human karyotype which comprises the steps: (a) contacting a preparation of said chromosomes, in single-stranded form, under conditions sufficient to permit nucleic acid hybridization to occur with a set of combinatorially labeled oligonucleotide probes, each member thereof: (i) having a predetermined fluorescent label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype; said set having sufficient members to be capable of specifically hybridizing each autosomal and sex chromosome of said human karyotype; wherein said contacting thereby results in each autosomal or sex chromosome of said preparation being hybridized to at least one member of said set of probes; (b) for each chromosome of said preparation hybridized to at least one member of said set of probes, employing an interferometer to detect and identify the predetermined label of the hybridized probe member(s) to correlate the identity of the label with the identity of the autosomal and sex chromosomes of said human karyotype, thereby identifying the chromosome hybridized to said probe member(s); and (c) repeating step (b) until each autosomal and sex chromosome of said human karyotype has been identified in said preparation.