Spatial positioning of spectrally labeled beads
Devices, systems, kits, and methods for detecting and/or identifying a plurality of spectrally labeled bodies well-suited for performing multiplexed assays. By spectrally labeling the beads with materials which generate identifiable spectra, a plurality of beads may be identified within the fluid. Reading of the beads is facilitated by restraining the beads in arrays, and/or using a focused laser.
1. A method comprising:
restraining a plurality of spectrally labeled bodies with affinity binding so as to define a multiplexed array, wherein the spectrally labeled bodies are spatially resolved and comprise a plurality of fluorescent labels used to encode discrete and different emission spectra wherein each emission spectrum has a plurality of signals at differing wavelengths;
directing an image of the multiplexed array of bodies onto a sensor to sense spectra generated by the bodies; and
identifying the bodies from the spectra sensed by the sensor.
2. The method of claim 1 , wherein the bodies are affixed to each other.
3. The method of claim 1 , wherein the bodies are restrained within an array of openings affixed in a support structure.
4. The method of claim 1 , wherein the sensor is a CCD camera.
5. A method of claim 1 , wherein the spectrally labeled bodies include polymers.
6. A method of claim 1 , wherein affinity binding is facilitated by a chemical moiety.
7. A method of claim 1 , wherein the affinity binding is specific.
8. A method of claim 1 , wherein the plurality of signals include a reference signal.
9. A method of claim 1 , wherein the affinity binding is facilitated with a binding pair.
10. The method of claim 1 , wherein the bodies are restrained to a support structure.
11. The method of claim 1 , wherein the bodies are restrained in the multiplexed array by an array of discrete binding sites.
12. A method of claim 8 , wherein the reference signal identifies a spectral code family.
13. A method of claim 9 , wherein one member of the binding pair is affixed to at least one of the plurality of spectrally labeled bodies and the other member is affixed to a site of the multiplexed array.
14. The method of claim 10 , wherein the support structure has a substantially planar surface.
15. The method of claim 10 , wherein the support structure contains a plurality of openings sized to accommodate one of the spectrally labeled bodies.
16. The method of claim 10 , wherein a plurality of unbound bodies are washed away.
17. A method of claim 11 , wherein the discrete binding sites are arranged so as to inhibit a presence of more than a single body at each binding site.
18. A method of claim 11 , wherein the binding sites comprise a material capable of binding to the bodies.
19. A method of claim 18 , wherein the material comprises a selective affinity molecule which selectively binds to one or more particular bodies.
20. The method of claim 19 , wherein the selective affinity molecule is streptavidin.
21. The method of claim 19 , wherein a selective binding material is a biotinylated structure.