( fect of optics misalignment on reporter...)Correlation of Bio-Plex Suspension Array Reader Validation With Multiplex Cytokine Assay Performance, Rev A

fect of optics misalignment on reporter channel performance, as shown in Table 3. In each case where the optics validation test fell outside acceptable specifications, certain parameters of the reporter validation test also showed unacceptable results. The slope decreased when the reporter %CV increased to 16%. When the slope decreased, the dynamic range also decreased. Sensitivity was significantly reduced at a reporter %CV of 16%. The accuracy was affected most significantly, with a value outside of acceptable specifications when the reporter %CV was >10.5%. These data indicate a high correlation between optics alignment and reporter channel performance.

Effect of Misaligned Optics on Cytokine Assay Performance
The effect of misaligned optics was also analyzed using a 3-plex cytokine assay (IL-6, IL-8, and GM-CSF) on a system at varying degrees of misalignment. The same parameters measured in the reporter validation kit were also evaluated for actual assays, including sensitivity, slope, and dynamic range. More specifically, five parameters were evaluated: 1) limit of detection (LOD) as defined by 3 standard deviations (SD) above the mean of the background, 2) low fluorescence intensity signal (equal to background), 3) high fluorescence intensity signal (10,000 pg/ml standard concentration), 4) dynamic range of the standard curve, and 5) signal-to-noise ratio of the 3.9 pg/ml standard. The results of the analyses of the three cytokines are shown in Table 4. When the optics alignment and reporter validation values fell outside acceptable specifications, the assay parameters changed significantly (Table 3). The signal-to-noise ratio and dynamic range were affected, while the LOD rem
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