Claims
- 1. A method of reducing flow analysis error in at least one flow analysis assay on a flow analyzer, wherein said flow analysis assay is performed subsequent to at least one prior flow analysis assay performed on the same flow analyzer, comprising:a) passing a population of one or more subsets of spectrally addressable microspheres, which had been exposed to a sample suspected of harboring one or more analytes of interest, through a flow analyzer, each subset of spectrally addressable microspheres being distinguishable by at least one characteristic from at least one other subset of spectrally addressable microspheres, said microspheres exhibiting a surface fluorescence value indicative of a biomolecular reaction having taken place on the surface of said microspheres in the presence of said one or more analytes of interest; b) discarding a number, N, of highest surface fluorescence values and a number, N, of lowest surface fluorescence values, which number equals a maximum number of microspheres, N, carried over from said prior assay performed on said same flow analyzer; and c) calculating the extent of the biomolecular reaction, if any, using surface fluorescence values not discarded in step b).
- 2. The method of claim 1 which further comprises determining coefficients of variation, mean, and standard deviation of the surface fluorescence values not discarded in step b).
- 3. The method of claim 2 in which the calculation of the extent of the biomolecular reaction is carried out only on those surface fluorescence values not discarded in step b that are within two standard deviations of the mean.
- 4. The method of claim 1 in which N is predetermined or estimated by a method comprising:a. passing a first sample comprising a known quantity of microspheres through a flow analyzer; b. passing a second sample comprising no microspheres through said flow analyzer; and c. calculating the number of microspheres detected by the flow analyzer in said second sample, wherein N, the maximum number of microspheres carried over from a prior assay, is the number of microspheres detected in the second sample.
- 5. The method of claim 4 in which a minimum number, M, of microspheres required to produce an assay having a low flow analysis error is estimated.
- 6. The method of claim 5 in which at least about 2N+M microspheres are passed through the flow analyzer.
- 7. The method of claim 1 which comprises a multiplexed assay.
CROSS-REFERENCE TO RELATED APPLICATION
The present application claims the benefit of the priority date of co-pending provisional patent application Ser. No. 60/149,226, filed Aug. 17, 1999, the complete disclosure of which is incorporated by reference herein.
US Referenced Citations (5)
Provisional Applications (1)
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Number |
Date |
Country |
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60/149226 |
Aug 1999 |
US |