Abstract

Photobleaching of fluorescent proteins prevents proper measurements and efficient image acquisition. For instance, the ratio measurements in FRET imaging are severely affected by photobleaching because donor and accepter molecules bleach at different rates [1]. The mechanism of photobleaching has been explored by a number of researchers [2]. In two photon excitation (TPE), the broad spectrum of an excitation laser such as Ti:S laser causes excited state absorption (ESA), which may trigger photobleaching.

© 2015 Japan Society of Applied Physics, Optical Society of America