Fixed polarizer ellipsometry for simple and sensitive detection of thin films generated by specific molecular interactions: applications in immunoassays and DNA sequence detection

• Published in: Clinical chemistry (Baltimore, Md.) Md.), 1998-09, Vol.44 (9), p.2031-2035
• Main Authors: Ostroff, Rachel M, Maul, Diana, Bogart, Gregory R, Yang, Shao, Christian, Jennifer, Hopkins, Deborah, Clark, David, Trotter, Brian, Moddel, Garret
• Format: Article
• Language: English
• Subjects: alpha-Fetoproteins - analysis; Antigens, Bacterial - analysis; Biological and medical sciences; DNA - chemistry; Fundamental and applied biological sciences. Psychology; General aspects; Hepatitis B Surface Antigens - blood; Humans; Immunoassay - instrumentation; Immunoassay - methods; Instrumentation. Materials. Reagents. Research laboratory organization; Light; Models, Theoretical; Nucleic Acid Hybridization - methods; Sensitivity and Specificity; Streptococcus pyogenes - immunology
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1093/clinchem/44.9.2031

Biological thin films may form on a surface by specific molecular interactions. The fixed polarizer ellipsometer (FPE) is a sensitive instrument that detects biological thin films either qualitatively or quantitatively. The design is simple and inexpensive. The assays are formatted on an optical surface, and the FPE detection is based on the phase shift of linearly polarized light after reflection through a thin film. We have constructed mathematical models of the FPE response to reflection through single-layer and two-layer films that agree closely with experimental data. Several biological assays have been measured with the FPE to demonstrate the application of this technology to clinical targets, including ultrasensitive immunoassays for hepatitis B surface antigen (0.1 ng/mL) and alpha-fetoprotein (0.01 ng/ mL) and DNA hybridization (0.5 fmol/microL target probe). A clinical study for detection of group A streptococcus from patient throat swabs demonstrated the qualitative application of the FPE to infectious disease targets. The flexibility and sensitivity of the FPE makes this technology suitable for numerous target analytes and applications.