Nanohole sensor chip with reference sections

A method for quantifying analytes using a plasmonic nanohole sensor chip which includes a substrate defining a plurality of nanoholes and a metal film disposed on the substrate, the sensor chip comprising a test section, upon which capture agents for the analyte are immobilized, and at least three reference sections, upon which capture agents conjugated with at least three different known quantities of the analyte are respectively immobilized, the method comprising:

• adding a sample to the test section of the sensor chip;
• applying a light to the sensor chip;
• comparing intensities of light transmitted through the test section and the reference sections of the sensor chip;
• and
• determining a presence and amount of the analyte in the sample based on the comparison.

A plasmonic nanohole sensor chip for detecting and assessing a quantity of an analyte in a test sample, comprising:

• a substrate, the substrate defining a plurality of nanoholes arranged in a periodic pattern;
• a metal film disposed on the substrate;
• an adhesion layer between the metal film and the substrate;
• capture agents for the analyte immobilized on a first section of the metal film;
• and
• a dielectric layer or an ion solution placed on a second section of the metal film, the second section including at least three reference sections, wherein in different ones of the at least three reference sections, the dielectric layer has different densities or the ion solution has different concentrations.