High-pulse-energy multiphoton imaging of neurons and oligodendrocytes in deep murine brain with a fiber laser
| Autor: | Hyungsik Lim, Michael J. Redlich, Brad Prall, Edesly Canto-Said, Yevgeniy Busarov, Lilit Shirinyan-Tuka, Arafat Meah |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Ytterbium Materials science Science chemistry.chemical_element Mice Transgenic Imaging techniques Article Green fluorescent protein law.invention 03 medical and health sciences 0302 clinical medicine law In vivo Fiber laser Animals Author Correction Neurons Multidisciplinary Lasers Brain Laser Fluorescence Cellular neuroscience Mice Inbred C57BL Oligodendroglia 030104 developmental biology Microscopy Fluorescence Multiphoton chemistry Sapphire Biophysics Medicine Biomedical engineering 030217 neurology & neurosurgery Ex vivo |
| Zdroj: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-11 (2021) |
| ISSN: | 2045-2322 |
| Popis: | Here we demonstrate high-pulse-energy multiphoton microscopy (MPM) for intravital imaging of neurons and oligodendrocytes in the murine brain. Pulses with an order of magnitude higher energy (~ 10 nJ) were employed from a ytterbium doped fiber laser source at a 1-MHz repetition rate, as compared to the standard 80-MHz Ti:Sapphire laser. Intravital imaging was performed on mice expressing common fluorescent proteins, including green (GFP) and yellow fluorescent proteins (YFP), and TagRFPt. One fifth of the average power could be used for superior depths of MPM imaging, as compared to the Ti:Sapphire laser: A depth of ~ 860 µm was obtained by imaging the Thy1-YFP brain in vivo with 6.5 mW, and cortical myelin as deep as 400 µm ex vivo by intrinsic third-harmonic generation using 50 mW. The substantially higher pulse energy enables novel regimes of photophysics to be exploited for microscopic imaging. The limitation from higher order phototoxicity is also discussed. |
| Databáze: | OpenAIRE |
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