1：Experimental Design and Method Selection:

The study involves designing a biosensor with super-hydrophobic plasmonic devices, utilizing metal enhanced fluorescence (MEF) for signal amplification. Methods include finite element methods (FEM) for electromagnetic field simulation, photochemical immobilization technique (PIT) for antibody functionalization, and fluorescence microscopy for detection.

2：Sample Selection and Data Sources:

Urine samples from healthy subjects and serum samples with known hematocrit values were used. Samples were selected from existing lab collections, including normal urine and those with potential interfering compounds like hemoglobin and bilirubin.

3：List of Experimental Equipment and Materials:

Equipment includes SEM (FEI Nova 600 NanoLab), AFM (ICON), fluorescence microscope (Leica DM 2700M), UV lamp (HERAEUS amalgam type NNI 40/20), fluidic circuits, and nephelometer (Siemens Healthcare Diagnostics BN II System). Materials include silicon wafers, S1813 positive tone resist, gold nanoparticles, antibodies (anti-human IgG), and FITC-tagged secondary antibodies.

4：Experimental Procedures and Operational Workflow:

Fabrication involved optical lithography and deep reactive ion etching to create micropillars decorated with gold nanoparticles. Functionalization used PIT with UV irradiation. Detection involved sandwich immunoassay with fluorescence measurement. Validation compared biosensor results with nephelometer measurements.

5：Data Analysis Methods:

Data were analyzed using linear regression for calibration curves, FEM simulations for EM field distribution, and statistical methods for confidence intervals and limits of detection.