TIRF is a fluorescent based detection method. Therein, laser light is coupled into a glass or polymer slide. Light is guided via total internal reflection and at each reflection point, an evanescent field is caused. The surface of the chip is covered by a sensitive detection layer and used recognition elements (e.g. antibodies) carry a fluorescent label which is excited by the evanescent field in case of binding to the surface.
As a labeled detection method, TIRF enables for
- Detecting very low concentrations
- Measuring in complex matrices (e.g. human serum)
Due to these features, TIRF found application in the area of water analysis and clinical diagnostics.
Literature
- Albrecht et al., Biosens. Bioelectron. 2010, 25(10), 2302-2308
- Barzen et al., Biosens. Bioelectron. 2002, 17(4), 289-295
- Kronick et al., J. Immun. Meth. 1975, 8(3), 235-240
