| RRC ID |
66017
|
| 著者 |
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.
|
| タイトル |
Dual-color deep-tissue three-photon microscopy with a multiband infrared laser.
|
| ジャーナル |
Light Sci Appl
|
| 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.
|
| 巻・号 |
7
|
| ページ |
12
|
| 公開日 |
2018-1-1
|
| DOI |
10.1038/s41377-018-0012-2
|
| PII |
12
|
| PMID |
30839589
|
| PMC |
PMC6107000
|
| IF |
14.0
|
| リソース情報 |
| ゼブラフィッシュ |
Tg(gfap:dTomato) |
