English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

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

MPS-Authors
/persons/resource/persons209303

Gregor,  C.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons41376

Steffens,  H.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

External Resource
No external resources are shared
Fulltext (public)

2486462.pdf
(Publisher version), 3MB

Supplementary Material (public)

2486462_Suppl_1.pdf
(Supplementary material), 4MB

2486462_Suppl_2.avi
(Supplementary material), 6MB

Citation

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.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002D-FF44-F
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.