total internal reflection fluorescence microscopy abbr., TIRFM (rus. микроскопия, флуоресцентная полного внутреннего отражения) — a technique for detection of fluorescent objects near interface using an optical microscope.


The total internal reflection fluorescence microscopy (TIRFM) technique uses the reflection of electromagnetic waves from the interface between two transparent media. This reflection occurs only if light travels from a medium of higher index of refraction at an angle larger than the critical angle (1/n). The intensity of light that enters the second medium decreases exponentially, which allows the detection of fluorescent objects excited by the light in the interface layer of ~100 nm thick at a resolution up to 10 nm [1]. Therefore, TIRFM may legitimately be referred to as one of the methods of fluorescence nanoscopy. In biological applications, this technique is used for imaging of the cell membrane and structures near the cell membrane.


  • Borisenko Grigory G.


  1. Saffarian S., Kirchhausen T. Differential evanescence nanometry: live-cell fluorescence measurements with 10-nm axial resolution on the plasma membrane // Biophys J. 2008. V. 94. P. 2333–2342.
  2. Truskey, G.A., Burmeister, J.S., Grapa, E. el al. Journal of Cell Science Volume 103, Issue 2, 1992, P. 491–499. Axelrod, D. Journal of Biomedical Optics Volume 6, Issue 1, January 2001, Pages 6–13.

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