Appeal No. 1997-2513
Application No. 08/206,917
Claims1 1 and 5 are illustrative of the subject matter on appeal and are
reproduced below:
1. A method for determining instrument event-counting linearity of a flow
cytometer comprising the steps of:
(a) separating a sample of cells into at least two aliquots;
(b) adding equal volumes of mixtures containing known concentrations of
fluorescent microparticles in a diluent to each aliquot, wherein the
concentration of microparticles added to each aliquot differs;
(c) counting the number of microparticles added to each aliquot by
means of flow cytometry; and
(d) performing statistical analysis on the number of microparticles
counted from all aliquots to determine linearity.
5. In a method for absolute counting of cells in a sample wherein the sample
comprises cells mixed with one or more cell markers having an emission
spectra and a known number of a first fluorescent microparticle having an
emission spectra, wherein the emission spectra of the cell markers and
the first microparticle are distinguishable, a method comprising the steps
of:
(a) separating the sample into at least two aliquots;
(b) adding equal volumes of mixtures containing known concentrations of
a second fluorescent microparticle in a diluent to each of the aliquots,
wherein the concentration of second fluorescent microparticles added
to each aliquot differs and wherein the second microparticle has an
emission spectra which is distinguishable from the emission spectra
of the first microparticle and cell markers;
(c) counting the number of fluorescent cells, the number of first
microparticles and the number of second microparticles in each
aliquot by means of flow cytometry; and
(d) performing statistical analysis on the number of cells counted, the
number of first microparticles counted and the number of second
microparticles counted in all aliquots to determine linearity.
Claim 12 is drawn to the method of claim 5, wherein the cell markers
comprise monoclonal antibodies labeled with carbocyanine, a fluorescent dye.
1 We note that the numbering of claims 7-11 as listed in the Appendix of appellants’
Brief is incorrect. Claim 7 has been canceled. Therefore, claims 7-11 should be
numbered 8-12, respectively.
2
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