Multiphoton microscopy allows allows imaging at much greater depth than standard single-photon fluorescence imaging. It is the gold-standard for imaging in thick tissues. The Olympus/Evident MPE comes with two broad-spectrum femtosecond pulsed lasers, a piezo stage, and a resonant scanner, making it ideal for imaging multiple fluorophores at high speed. It also has an additional scanner for simultaneous photomanipulation of the smaple while imaging.

The system comes with fluorescence lifetime imaging (FLIM) capabilities from PicoQuant for quantitative biosensing experiments. The MPE is fully motorised and has an incubator, making it excellent for imaging live samples, in particular embryos and tissues.

Located in the basement of IBRB in room IBRB03.

Hardware details for methods reporting in publications.

Filter diagram showing available options.

Absorption spectra of some dyes (no promises on the reliability of these papers):

https://doi.org/10.1046/j.1365-2818.2002.01074.x

https://doi.org/10.1111/jmi.12255

https://doi.org/10.3390%2Fijms232113407

https://doi.org/10.1016/j.bpj.2011.12.056

Excel table from UofRochester

See fpbase.org for other dyes and fluorescent proteins.

Booking rules for MPE.

Does using the fast resonant scanner help with photo-bleaching/toxicity? This seems unclear, and the best approach is to test with your sample. See the following for discussions on the microlist forum, Peter Rupprecht's blog, and also a paper comparing photobleaching for one and two-photon exictation.

The .oir files open in Fiji (not ImageJ) by just drag dropping, but sometimes there are problems and you have to install a separate plugin, see here for instructions.

Please acknowledge the below funding in your publications, in addition to following the usual CAMDU acknowledgment policy.

BBSRC BB/T018070/1