Devices, systems, and methods for conducting assays with improved sensitivity using sedimentation
View Patent ↗Embodiments of the present invention are directed toward devices, systems, and method for conducting assays using sedimentation. In one example, a method includes layering a mixture on a density medium, subjecting sedimentation particles in the mixture to sedimentation forces to cause the sedimentation particles to move to a detection area through a density medium, and detecting a target analyte in a detection region of the sedimentation channel. In some examples, the sedimentation particles and labeling agent may have like charges to reduce non-specific binding of labeling agent and sedimentation particles. In some examples, the density medium is provided with a separation layer for stabilizing the assay during storage and operation. In some examples, the sedimentation channel may be provided with a generally flat sedimentation chamber for dispersing the particle pellet over a larger surface area.
1. A method for conducting an assay comprising:
layering a mixture on a density medium in an assay area, wherein the mixture includes a sample, a first separation layer fluid, and sedimentation particles, wherein the sedimentation particles have a density greater than the density medium, and wherein said layering a mixture comprises forming, with the first separation layer fluid, an interface between the density medium and the sample, between the sample and the sedimentation particles, or between the density medium and the sedimentation particles;
subjecting the mixture to sedimentation forces such that the sedimentation particles, if formed, travel through the first separation layer fluid and the density medium to a detection area; and
detecting a presence of an analyte of interest in the detection area.
2. The method of claim 1 , wherein the first separation layer fluid comprises a material which is denser than the density medium but less dense than the coated particles and wherein said layering a mixture further comprises positioning the material near the detection area.
3. The method of claim 1 , wherein the first separation layer fluid and density medium are isolated by hydrophobic interaction.
4. The method of claim 1 , wherein the first separation layer fluid and density medium are isolated by high viscosity.
5. The method of claim 1 , wherein the first separation layer fluid and density medium are isolated by charged or water ordering polymer and/or salt interaction.
6. The method of claim 1 , wherein the first separation layer fluid comprises a material which is denser than the sample but less dense than the density medium and wherein said layering a mixture further comprises forming the interface between the density medium and the sample.
7. The method of claim 6 , wherein said subjecting the mixture further comprises preventing at least a portion of free labeling agent provided in the sample from being transported through the density medium.
8. The method of claim 1 , wherein the sedimentation particles include particles having a negatively charged surface.
9. The method of claim 1 , wherein said subjecting the mixture comprises spinning a substrate, and wherein the substrate at least partially supports the mixture.
10. The method of claim 1 , wherein the detection area has a first thickness smaller than a thickness of a microfluidic channel containing the density medium.
11. The method of claim 1 , further comprising:
collecting the sedimentation particles at an end portion of a microfluidic channel, the end portion being narrower than a main portion of the microfluidic channel, containing the density medium.
12. The method of claim 1 , wherein the first separation layer fluid comprises a hydrophobic material, a mineral oil, an organic oil, a charged or water ordering polymer, a salt in a water based medium, a fluoroalkane fluid, a perfluorocarbon, or a perfluoroalkane fluid.
13. The method of claim 1 , wherein the assay area is further provided with a second separation layer.
14. The method of claim 13 , wherein the first separation layer fluid comprises providing a first material which is denser than the sample but less dense than the density medium, wherein the second separation layer comprises a second material which is denser than the density medium but less dense than the coated particles, and
wherein said layering a mixture further comprises positioning the first material between the density medium and the sample and positioning the second material near the detection area.
15. The method of claim 1 , wherein the density media is a liquid having a selected density.