IGB Core Instruments Zeiss LSM 710

Light Path

https://www.gu.se/en/core-facilities/lsm-710-nlo

Seven visible excitation lines: 405nm, 458nm, 488nm, 514nm, 561nm, 594nm, 633nm.

Tisaphire laser 700nm to 980nm

 

Spectral

 

Unmixing

Two Photon: Image Deeper with Less Photo Damage

Two photon uses two photons to excite a single electronic transition. this means that both photons need to be absorbed simultaneously.  Single photon absorption follows Beers law

http://zeiss-campus.magnet.fsu.edu/articles/spectralimaging/introduction.html

Multiphoton Microscopy https:http://en.wikipedia.org/wiki/Two-photon_absorption#Absorption_ratezeiss-campus.magnet.fsu.edu/referencelibrary/multiphoton.html 

Fluorescence Lifetime Imaging Microscopy (FLIM): Separate fluorophores based on average time in the excited state.

start with single photon counting and decay curveshttp://www.iss.com/microscopy/components/FastFLIM.html

 

then introduce Phasor plots