Composition, synthesis and use of simulated cells

Claim 1

A method of determining the effective fluorescence F/P ratio of a fluorescent antibody, comprising: (a) providing first microbeads having binding sites capable of binding to the fluorescent antibodies, said microbeads being of known or determinable number of binding sites per microbead; (b) providing second calibration microbeads having associated therewith the same fluorescent dye as said fluorescent antibodies, and exhibiting the same fluorescence spectra as said fluorescent antibodies; (c) providing a flow cytometer; (d) establising a calibration plot for the flow cytometer with said second calibration microbeads, of number of equivalent soluble fluorescent dye molecules per microbead as a function of fluorescence intensity channel of said flow cytometer; (e) reacting said first microbeads with excess fluorescent antibodies, to yield fluorescent antibody bound microbeads; (f) determining the flourescence intensity channel of said fluorescent antibody-bound microbeads on said flow cytometer; (g) for said fluorescence intensity channel determined for said fluorescent antibody-bound microbeads, determining from said calibration plot the number of equivalent soluble fluorescent dye molecules per microbead for said fluorescent antibody-bound microbeads; and (h) dividing the number of equivalent soluble fluorescent dye molecules per microbead determined in step (f) by the number of binding sites per microbead, to yield said effective fluorescence F/P ratio of said fluorescent antibody.

Claim 2

A method of determining the effective fluorescence F/P ratio of a fluorescent antibody, comprising: (a) providing first microbeads having binding sites capable of binding to the fluorescent antibodies, said microbeads being of known or determinable number of binding sites per microbead; (b) providing second calibration microbeads having associated therewith the same fluorescent dye as said fluorescent antibodies, and exhibiting the same fluorescence spectra as said fluorescent antibodies; (c) providing a flow cytometer; (d) establising a calibration plot for the flow cytometer with said second calibration microbeads, of number of equivalent soluble fluorescent dye molecules per microbead as a function of fluorescence intensity channel of said flow cytometer; (e) reacting said first microbeads with excess fluorescent antibodies, to yield fluorescent antibody bound microbeads; (f) determining the flourescence intensity channel of said fluorescent antibody-bound microbeads on said flow cytometer; (g) for said fluorescence intensity channel determined for said fluorescent antibody-bound microbeads, determining from said calibration plot the number of equivalent soluble fluorescent dye molecules per microbead for said fluorescent antibody-bound microbeads; and (h) dividing the number of equivalent soluble fluorescent dye molecules per microbead determined in step (f) by the number of binding sites per microbead, to yield said effective fluorescence F/P ratio of said fluorescent antibody.