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  In vivo mouse and live cell STED microscopy of neuronal actin plasticity using far-red emitting fluorescent proteins.

Wegner, W., Ilgen, P., Gregor, C., van Dort, J., Mott, A. C., Steffens, H., et al. (2017). In vivo mouse and live cell STED microscopy of neuronal actin plasticity using far-red emitting fluorescent proteins. Scientific Reports, 7: 11781. doi:10.1038/s41598-017-11827-4.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-FF44-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-FF49-5
Genre: Journal Article

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 Creators:
Wegner, W., Author
Ilgen, P., Author
Gregor, C.1, Author              
van Dort, J., Author
Mott, A. C., Author
Steffens, H.1, Author              
Willig, K. I., Author
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1Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, ou_578627              

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 Abstract: The study of proteins in dendritic processes within the living brain is mainly hampered by the diffraction limit of light. STED microscopy is so far the only far-field light microscopy technique to overcome the diffraction limit and resolve dendritic spine plasticity at superresolution (nanoscopy) in the living mouse. After having tested several far-red fluorescent proteins in cell culture we report here STED microscopy of the far-red fluorescent protein mNeptune2, which showed best results for our application to superresolve actin filaments at a resolution of similar to 80 nm, and to observe morphological changes of actin in the cortex of a living mouse. We illustrate in vivo far-red neuronal actin imaging in the living mouse brain with superresolution for time periods of up to one hour. Actin was visualized by fusing mNeptune2 to the actin labels Lifeact or Actin-Chromobody. We evaluated the concentration dependent influence of both actin labels on the appearance of dendritic spines; spine number was significantly reduced at high expression levels whereas spine morphology was normal at low expression.

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Language(s): eng - English
 Dates: 2017-09-18
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/s41598-017-11827-4
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Title: Scientific Reports
Source Genre: Journal
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Pages: 10 Volume / Issue: 7 Sequence Number: 11781 Start / End Page: - Identifier: -