Using single molecule microscopy technique to achieve elongated time-period blinking using total internal reflection technique

Abstract EN

Recent advances in field of optical microscopy have made it possible to detect image and obtain spectroscopic data from individual molecules, revealing information about population distributions (both static and dynamic) as well as photophysical information.

As higher levels of time resolution become available layers of dynamic information are uncovered, elucidating the fine detail of chemical reaction normally hidden within the ensemble.

Single molecule (SM) techniques are especially relevant to macromolecular biochemistry as chemical processes occurring within large molecules are often too complicated to study in the ensemble.

Single Molecule Fluorescence Microscopy (SMFM) is achieved using coherent laser light to excite the fluorophore to a singlet excited state, from which it can relax rapidly.

However this can be followed by either absorption of additional photons, eventually lead to an ionised or “photo-bleached” state, or spin-inversion and non-radiative intersystem crossing to a triplet state.

The excitation volume has been reduced to achieve elongated time-period blinking using Total Internal Reflection.