Digital LSPR for enhanced assay sensitivity
Systems, methods, and devices related to detecting a presence of an analyte and/or determining a concentration of analytes are provided. An analyte may be provided on an LSPR-active surface. The LSPR-active surface may comprise sensitivity enhancing labels. The analyte may induce a local change near the LSPR-active surface. The LSPR-active surface may be imaged with an imaging device for images before, during, or after a reaction takes place. Local regions of interest within the images may be analyzed to detect the local changes.
1. A computer-implemented method, comprising:
randomly selecting a plurality of corresponding regions of interest in each of a series of two or more images of a nanostructured sensor surface, wherein the nanostructured sensor surface is capable of sustaining a localized surface plasmon resonance;
measuring a change in RGB value or greyscale value for each of the plurality of corresponding regions by comparing the RGB value or greyscale value for corresponding regions of interest in different images of the series of two or more images; and
determining a presence of an analyte based on analyzing the measured changes in RGB value or greyscale value for the plurality of corresponding regions.
2. The method of claim 1 , wherein a limit of detection for determining the presence of the analyte is determined to be better than 1 ng/mL based on analyzing a control reading and one or more readings corresponding to one or more known quantities or concentations of the analyte.
3. The method of claim 1 , wherein a limit of detection for determining the presence of the analyte is determined to be better than 1 pg/mL based on analyzing a control reading and one or more readings corresponding to one or more known quantities or concentations of the analyte.
4. The method of claim 1 , further comprising determining a concentration of the analyte based on the measured changes in RGB value or greyscale value.
5. The method of claim 1 , wherein the measured change in RGB value or greyscale value is a change in RGB value or greyscale value of light reflected from the nanostructured sensor surface.
6. The method of claim 1 , wherein the series of two or more images are captured before and after a local analyte-induced change occurs.
7. The method of claim 1 , wherein the nanostructured sensor surface is contacted with a primary binding component, the analyte, and a secondary binding component sequentially or simultaneously, wherein the secondary binding component is a sensitivity enhancing label.
8. The method of claim 7 , wherein the sensitivity enhancing label is an enzyme that catalyzes a conversion of a reactant to an insoluble product, or a metallic nanoparticle that is capable of inducing plasmon-plasmon coupling between the metallic nanoparticle and the nanostructured sensor surface.
9. The method of claim 1 , wherein the analyte is present in a sample an amount of 100 ng/mL or less.
10. The method of claim 1 , wherein the analyte is present in a sample in an amount of 1 pg/mL or less.
11. The method of claim 1 , further comprising receiving a report comprising a result of the method and making a healthcare decision based on the reported result, wherein the sample is a patient sample.
12. The method of claim 1 , wherein the nanostructured sensor surface comprises a continuous nanostructured metallic thin film across a sensor substrate.
13. The method of claim 1 , wherein determining the presence of the analyte comprises:
analyzing measured changes in RGB value and intensity of light for the plurality of corresponding regions.
14. The method of claim 13 , wherein the measured changes in RGB value and intensity of light for the plurality of corresponding regions are based on comparing the RGB value and intensity of light for corresponding regions of interest in different images of the series of two or more images.
15. The method of claim 1 , further comprising:
generating a histogram relating to the measured changes in RGB value or greyscale value for the plurality of corresponding regions, wherein the determining the presence of the analyte is based on the histogram.
16. The method of claim 1 , wherein analyzing the measured changes in RGB value or greyscale value for the plurality of corresponding regions comprises:
identifying one or more statistically significant differences in the measured changes in RGB value or greyscale value.
17. The method of claim 1 , wherein the nanostructured sensor surface comprises: a first metallic thin film layer, a second metallic thin film layer, and a dielectric layer at least partially between the first metallic thin film layer and the second metallic thin film layer.
18. The method of claim 1 , wherein the nanostructured sensor surface comprises: a first plurality of metallic thin film layers, a second plurality of metallic thin film layers, and a dielectric layer at least partially between the first plurality of metallic thin film layers and the second plurality of metallic thin film layers.