Method for accurate counting of probe spots in cell nuclei

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 911, 435 912, 436501, 436 63, 436 94, 536 243, 536 2432, 2504621, 549223, 935 76, 935 77, 935 78, C12Q 168, F21K 200, C07H 2104, C12N 1500

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055232078

ABSTRACT:
A method for the accurate counting of DNA probe spots in cell nuclei wherein anomalies caused by a two dimensional measurement of a three dimensional cell sample are eliminated from evaluation. DNA probe spots in cell nuclei which are counted by means of Fluorescent In Situ Hybridization (FISH) include cells wherein probe spots of different contoured cells are overlaid or are detected as being adjacent one another with resultant erroneous diagnostic results such as with cancer detection or prognosis. A gating fluorescent value is determined by clusters of fluorescence in regions of non-anomalous values of fluorescence determined by plotting peak fluorescent value against area. The loci of the non-anomalous peak values cluster in specifically definable regions whereby fluorescent values for cells which deviate from the gating fluorescent value, are discounted in the preparation of histograms or other diagnostic measurements.

REFERENCES:
patent: 5072382 (1991-12-01), Kamentsky
Martin-Reay et al., American Journal of Clinical Pathology, vol. 102, No. 4, Oct. 1991, pp. 432-438.
Lichter et al., GATA 8(1):24-35, 1991.

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