The bh SPC-150 TCSPC FLIM modules have been integrated in the STED FLIM microscopes of Abberior Instruments GmbH, Göttingen, Germany. The system is based on Stefan Hell’s stimulated-emission-depletion technique, see ‘Nanoscopy with Focused Light (Nobel Lecture)’, Angewandte Chemie, June 2015. The system records single STED FLIM images or 3D stacks of STED FLIM images. In single images the fluorescence intensity and lifetime are mapped at a lateral resolution of < 30 nm. 3D FLIM stacks are recorded at a longitudinal and lateral resolution of <70nm. Four FLIM detection channels are available; three-channel STED measurements are performed with a single depletion laser. The system is able to separate the signals of several fluorophores by their fluorescence lifetimes, or to use the fluorescence lifetime as a probe function for the molecular environment. Moreover, variable post-process time-gating allows the user to tune the optical resolution of the images versus the signal level.
For more information please see bh TCSPC Handbook and abberior-instruments.com/products/superresolution-microscopes.
Confocal (left) and STED FLIM (right) image recorded with Abberior Instruments STED FLIM system. Mammalian cells labelled with tubulin/ Atto647N and vimentin/ Abberior STAR 635P.