Nanoarray-in-microarray multiplexed analysis methods and systems
AssigneeThe Royal Institution for the Advancement of Learning/Mcgill University
InventorsDavid Juncker, Gina Zhou, Sebastien Bergeron
Life of this patent
Estimating the term…
Abstract
Methods and apparatuses for performing a nanoarray-in-microarray assay is provided, which can be used to estimate a protein concentration in a sample solution. A plurality of nanodots are fabricated on a surface having at least one affinity binder. One or more microspots are superimposed over the nanodots on predetermined regions of the surface, each of the microspots comprising at least one antibody. An assay process is performed on the surface, and the surface is imaged to acquire optical images of the nanodots within each microspot. Image analysis algorithms are the performed on the optical images to identify bindings on individual ones of the plurality of nanodots.
Claims20
A method for performing an assay, comprising:
- fabricating a plurality of nanodots on a surface having at least one affinity binder;
- applying a microspot on a predetermined region of the surface to superimpose the microspot over a number of the nanodots to form a nanoarray-in-microarray having the number of the nanodots within a perimeter of the microspot, the microspot comprising at least one antibody of a plurality of antibodies;
- performing an affinity-binding assay process on the surface;
- acquiring at least one optical image of the number of nanodots within the microspot;
- and
- performing an image analysis on the at least one optical image to identify bindings on individual ones of the number of nanodots.
An assay device, comprising;
- a detection surface having defined thereon a plurality of nanodots each having at least one affinity binder;
- and
- a microspot applied on a predetermined region of the detection surface such that the microspot is superimposed over a number of the nanodots to form a nanoarray-in-microarray having the number of the nanodots within a perimeter of the microspot, the microspot comprising at least one antibody of a plurality of antibodies.
A method for estimating a protein concentration in a sample solution, comprising:
- forming a nanoarray-in-microarray by immobilizing nanodots of capture antibodies mixed with fluorescently-labeled detection antibodies on a microarray slide using nano-contact printing to superimpose a nanoarray of the nanodots on a predetermined region of the microarray slide having microspots forming a microarray, wherein a number of the nanodots are disposed within a perimeter of one or more of the microspots of the microarray;
- performing a sandwich assay with the nanoarray-in-microarray by sequentially incubating the sample solution with the fluorescently-labelled detection antibodies;
- imaging results of the assay to acquire a plurality of two-colour fluorescent images;
- overlaying the acquired two-colour fluorescent images to reveal at least one co-localized spot;
- and
- performing an image analysis algorithm to count the at least one co-localized spot to estimate the protein concentration.
Dependencies are inferred from the claim text.