Module 3 : Microscopic techniques

Lecture 15 : Light Microscopy-II

Total internal reflection fluorescence (TIRF)

The phenomenon of total internal reflection is described in lecture 10 (Figure 10.5). The evanescent field decays exponentially; the molecules in the close proximity of the slide/culture plate are selectively excited. The fluorescence therefore, is observed from a thin layer of the sample. Such an arrangement is particularly useful for studying membrane proteins (Figure 15.5).

Figure 15.5 A diagrammatic representation of studying membrane proteins using TIRF