Fig 1: Multiplex of the IL-6 QLISA (fluorescent signal from QdotTM 525) and the GFAP QLISA (fluorescent signal from QdotTM 585) where the sample consisted of 25,000 pg/mL IL-6 and 10,000 pg/mL GFAP. (A) Representative images of ~4.5 µm beads trapped in the IL-6 detection band of the variable height device (left) and ~2.8 µm beads trapped in the GFAP detection band (right). Each image is an overlay of the QdotTM 525 and QdotTM 585 channels, and the brightness/contrast was set for ease of viewing. (B) The fluorescence intensity values from the QdotTM 525 (blue) and QdotTM 585 (yellow) channels for each variable height device detection band shown in (A) with the control values subtracted. The error bars are standard error of the mean and represent the variability in fluorescence intensity between the fields of view within each detection band.
Fig 2: QLISA standard curves for GFAP in (A) spiked human serum and (B) spiked human whole blood, (C) IL-6 in spiked buffer, and (D) IL-8 in spiked buffer. Each data point represents three replicates (n = 3). (E) Comparison of the GFAP concentrations determined by the Quanterix Simoa® assay and by the QLISA method for ten clinical serum samples. The dashed red line represents the line y = x, and the solid black line represents the trend between the Quanterix GFAP concentration and the QLISA GFAP concentration. Each black dot represents a serum sample from a different donor. Replicates ranged from n = 1 to n = 5 depending on the volume of the serum sample. All error bars throughout the figure are standard error of the mean.
Fig 3: Multiplex of the IL-6 QLISA (fluorescent signal from QdotTM 525), the GFAP QLISA (fluorescent signal from QdotTM 585), and the IL-8 QLISA (fluorescent signal from QdotTM 655) where the sample consisted of 25,000 pg/mL IL-6, 10,000 pg/mL GFAP, and 1000 pg/mL IL-8. (A) Representative images of ~4.5 µm beads trapped in the IL-6 detection band of the variable height device (left), ~2.8 µm beads trapped in the GFAP detection band (center), and ~1 µm beads trapped in the IL-8 detection band (right). Each image is an overlay of the QdotTM 525, QdotTM 585, and QdotTM 655 channels. The brightness/contrast of each image was set for ease of viewing. (B) The fluorescence intensity values from the QdotTM 525 (blue), QdotTM 585 (yellow), and QdotTM 655 (magenta) channels for each variable height device detection band shown in (A) with the control values subtracted. The error bars are standard error of the mean and represent the variability in fluorescence intensity between the fields of view within each detection band.
Fig 4: Multiplex of the IL-6 QLISA and the GFAP QLISA where the fluorescent signal for both assays is from QdotTM 585. Both assays were run in the same microcentrifuge tube on the same spiked buffer sample. The representative fluorescence images from the variable height device detection bands of (A) a sample containing 25,000 pg/mL IL-6 and 10,000 pg/mL GFAP, (B) a sample containing 25,000 pg/mL IL-6 and 0 pg/mL GFAP, (C) a sample containing 0 pg/mL IL-6 and 10,000 pg/mL GFAP, and (D) a sample containing 0 pg/mL IL-6 and 0 pg/mL GFAP all have the same brightness/contrast for ease of visual comparison.
Supplier Page from Abcam for Human IL-6 Matched Antibody Pair Kit