PUBLICATION
Dual-color deep-tissue three-photon microscopy with a multiband infrared laser
- Authors
- Guesmi, K., Abdeladim, L., Tozer, S., Mahou, P., Kumamoto, T., Jurkus, K., Rigaud, P., Loulier, K., Dray, N., Georges, P., Hanna, M., Livet, J., Supatto, W., Beaurepaire, E., Druon, F.
- ID
- ZDB-PUB-190308-10
- Date
- 2018
- Source
- Light, science & applications 7: 12 (Journal)
- Registered Authors
- Dray, Nicolas
- Keywords
- none
- MeSH Terms
- none
- PubMed
- 30839589 Full text @ Light Sci Appl
Citation
Guesmi, K., Abdeladim, L., Tozer, S., Mahou, P., Kumamoto, T., Jurkus, K., Rigaud, P., Loulier, K., Dray, N., Georges, P., Hanna, M., Livet, J., Supatto, W., Beaurepaire, E., Druon, F. (2018) Dual-color deep-tissue three-photon microscopy with a multiband infrared laser. Light, science & applications. 7:12.
Abstract
Multiphoton microscopy combined with genetically encoded fluorescent indicators is a central tool in biology. Three-photon (3P) microscopy with excitation in the short-wavelength infrared (SWIR) water transparency bands at 1.3 and 1.7 µm opens up new opportunities for deep-tissue imaging. However, novel strategies are needed to enable in-depth multicolor fluorescence imaging and fully develop such an imaging approach. Here, we report on a novel multiband SWIR source that simultaneously emits ultrashort pulses at 1.3 and 1.7 µm that has characteristics optimized for 3P microscopy: sub-70 fs duration, 1.25 MHz repetition rate, and µJ-range pulse energy. In turn, we achieve simultaneous 3P excitation of green fluorescent protein (GFP) and red fluorescent proteins (mRFP, mCherry, tdTomato) along with third-harmonic generation. We demonstrate in-depth dual-color 3P imaging in a fixed mouse brain, chick embryo spinal cord, and live adult zebrafish brain, with an improved signal-to-background ratio compared to multicolor two-photon imaging. This development opens the way towards multiparametric imaging deep within scattering tissues.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping