Multiplexed assay systems and methods
A marker and sample technology, applied in measurement devices, biological testing, material analysis by observing the impact on chemical indicators, etc., can solve problems such as reducing the efficiency of handling droplets, limiting the efficiency of droplet analysis, and time-consuming
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example 1
[0111] Figures 14A-15B An exemplary use of marker particles with darkness shifting is illustrated. as above and Figure 14A As shown in , a marker particle 1410 can include a body 1412 (eg, a bead) and a capture material 1414 that includes a capture surface. Figure 14B Schematic diagram of the enzyme protocol on the capture surface utilizing the enzyme-linked darkening assay technique. in particular, Figure 14B Marker particles that can undergo a darkness shift via a "sandwich" arrangement are illustrated. In this sandwich arrangement, the analyte of interest 1440 is bound between two antibodies, including a capture antibody 1430 and an enzyme-conjugated detection antibody 1450 . When mixed with a suitable detection catalyst and probe 1470 with an enzyme substrate (detection substrate), the enzyme substrate is consumed by the enzyme on the primary antibody 1450, which results in darkened precipitation. This precipitation causes a color change on the capture surface (e...
example 2
[0115] Furthermore, as described above, the detection of different kinds of marker particles that undergo a darkness shift (e.g., differentiated based on different modalities) can facilitate the association of different analytes of interest with marker particles in a multiplexed fashion. analysis. E.g, Figure 19 The multiplexing application of darkness-shifted marker particles using three different marker particle types is illustrated. in particular, Figure 19 Application of size-based multiplexing of darkness-shifted marker particles is illustrated. The first marker particle type A includes first spherical beads with a size of about 10 μm, the second marker particle type B includes second spherical beads with a size of about 15 μm, and the third marker particle type C includes first spherical beads with a size of about 20 μm. Three spherical beads. The agglomeration process was used to coat the beads with hydrogel. The capture antibody is attached to the marker particl...
example 3
[0117]Another example of size-based multiplexing applications involves the use of monodisperse hydrogels of different sizes. Except that the first marker particle type A may comprise hydrogel spheres of a first size (eg, 5 μm), the second marker particle type B may comprise hydrogel spheres of a second size (eg, 10 μm) and a third marker particle type Example 3 can be similar to Example 2 above, except that particle type C can include hydrogel spheres of a third size (eg, 15 μm). As in Example 2, the marker particles darken upon visualization of their associated analyte and enzyme reactions (eg, in a sandwich ELISA). However, instead of differentiating between marker particle types based on internal bead size, in this example marker particle types can be differentiated based on overall marker particle size.
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