Brownian microbarcodes for bioassays
An encoded microparticle, methods for using the same in bioassays, and a method of making the same are provided herein.
1 . A method for detecting an analyte in a test fluid, comprising:
providing a set of biochemically active microparticles, each microparticle comprising a spatial code;
wherein a layer of the microparticles is arranged on a surface during analysis,
detecting electromagnetic radiation from the microparticles in order to detect the spatial codes of the individual microparticles; and
wherein the microparticles are configured such that they undergo substantial Brownian motion.
2 . The method of claim 1 , wherein the microparticles extend in a line or plane.
3 . The method of claim 1 , wherein the electromagnetic radiation is reflected light from the microparticles.
4 . The method of claim 1 , wherein the electromagnetic radiation is transmitted light through the microparticles.
5 . The method of claim 1 , wherein the electromagnetic radiation is luminescence optical emission from the microparticles.
6 . The method of claim 1 , wherein a two dimensional diffusion coefficient of the microparticles is greater than 1×10 −12 cm 2 /s.
7 . A method for testing a biochemical sample, comprising:
providing a set of biochemically active microparticles, each microparticle comprising a spatial code;
wherein a layer of the microparticles is arranged on a surface during analysis,
detecting electromagnetic radiation from the microparticles in order to detect the spatial codes of the individual microparticles; and
wherein the microparticles undergo a lateral displacement of 20 nm or greater in a time interval of one second or less.
8 . The method of claim 7 , wherein the microparticles extend in a line or plane.
9 . The method of claim 7 , wherein the electromagnetic radiation is reflected light from the microparticle.
10 . The method of claim 7 , wherein the electromagnetic radiation is transmitted light through the microparticle.
11 . The method of claim 7 , wherein the electromagnetic radiation is luminescent emission light from the microparticle.
12 . The method of claim 7 , wherein the two dimensional diffusion coefficient of the microparticles is greater than 1×10 −12 cm 2 /s.
13 . A container comprising:
a liquid;
a plurality of microparticles in the liquid, the microparticles having a biochemically active material on a surface; and
wherein the microparticles have a spatial code detectable under visible light;
wherein the microparticles undergo substantial Brownian motion in the liquid.
14 . The method of claim 13 , wherein the microparticles extend in a line or plane.
15 . The method of claim 13 , wherein a two dimensional diffusion coefficient of the microparticles is greater than 1×10 −12 cm 2 /s.
16 . A method of determining the result of a bioassay comprising:
providing a multiplicity of non-spherical encoded microparticles in a liquid;
quantitating optical signals from the microparticles;
wherein the microparticles are undergoing random molecular displacements during quantitation.
17 . The method of claim 16 , wherein the random molecular displacements for at least 10% of the microparticles exceed 20 nm or greater in a time interval of 5 seconds or less.
18 . The method of claim 17 , wherein the two dimensional diffusion coefficient of the microparticles is greater than 1×10 −12 cm 2 /s.
19 . A method for detecting the spatial codes of elongated microbarcodes, comprising:
providing the elongated microbarcodes with a biological material thereon;
binding the biological material on at least some of the microbarcodes with corresponding biological analyte in a test sample;
providing the elongated microbarcodes in a test fluid;
detecting electromagnetic radiation from the microbarcodes in order to detect the spatial codes of the individual microbarcodes;
detecting fluorescence on at least some of the microbarcodes in order to determine the presence of molecular binding events; and
wherein the microbarcodes undergo substantial Brownian motion in the test fluid during the detection of the spatial codes and during the detection of the molecular binding events.
20 . The method of claim 19 , wherein the two dimensional diffusion coefficient of the microbarcodes is greater than 1×10 −12 cm 2 /s.
21 . The method of claim 19 , wherein molecular binding events are indicated by fluorescence.
22 . The method of claim 19 , wherein more than 10% of the microbarcodes are measured to undergo a lateral displacement of 20 nm or greater in a time interval of one second or less.
23 . The method of claim 19 , wherein the microbarcodes further exhibit fluorescence as a measure of molecular binding events.
24 . The method of claim 19 , wherein the microbarcodes are omni-directional encoded particles with each microbarcode comprising a first material comprising two or more discrete segments aligned along an axis, and a second material surrounding the first material such that the segments are detectable through the second material, whereby a code is formed by the segments and the code is detectable from all directions perpendicular to the axis.
25 . The method of claim 19 , wherein the spatial codes are produced with projection photolithography.
26 . (canceled)
27 . The method of claim 19 , wherein the spatial codes are comprised of code elements that have a physical size less than 1 micron.
28 . (canceled)
29 . The method of claim 19 , wherein the biologically active material is selected from the group consisting of a nucleic acid, protein, antibody, enzyme, drug, receptor, and a ligand.
30 . The method of claim 19 , wherein the microbarcodes have a bar shape with an aspect ratio greater than 2:1.
31 . The method of claim 19 , wherein a cross-section taken orthogonal to the longest axis of the microbarcode is substantially square.
32 . The method of claim 19 , wherein the microbarcodes have a largest dimension of 50 microns.
33 . The method of claim 7 , wherein the surface is an inner surface of a container.
34 . The method of claim 33 , wherein the container is a microtiter plate.
35 . The method of claim 1 , wherein the surface is an inner surface of a container.
36 . The method of claim 35 , wherein the container is a microtiter plate.
37 . The method of claim 16 , wherein the bioassay comprises a nucleic acid, protein, peptide, polypeptide, polynucleotide, oligonucleotide, cell, antibody, enzyme, drug, receptor, ligand, or lipid.
38 . The method of claim 16 , wherein the microparticles are omni-directional encoded microparticles with each microparticle comprising a first material comprising two or more discrete segments aligned along an axis, and a second material surrounding the first material such that the segments are detectable through the second material, whereby a code is formed by the segments and the code is detectable from all directions perpendicular to the axis.
39 . The method of claim 16 , wherein the microparticles have a largest dimension of 50 microns.
40 . A biochemically active, non-spherical microparticle comprising a spatial code, and an elongated shape and is disposed in an environment such that the microparticle undergoes substantial Brownian motion in the liquid.