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  Nanoscopy of living brain slices with low light levels.

Testa, I., Urban, N., Jakobs, S., Eggeling, C., Willig, K. I., & Hell, S. W. (2012). Nanoscopy of living brain slices with low light levels. Neuron, 75(6), 992-1000. doi:10.1016/j.neuron.2012.07.028.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-EE48-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-1F73-4
Genre: Journal Article

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 Creators:
Testa, I.1, Author              
Urban, N.1, Author              
Jakobs, S.2, Author              
Eggeling, C.1, Author              
Willig, K. I.1, Author              
Hell, S. W.1, Author              
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              
2Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              

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 Abstract: Lens-based fluorescence microscopy, which has long been limited in resolution to about 200 nanometers by diffraction, is rapidly evolving into a nanoscale imaging technique. Here, we show that the superresolution fluorescence microscopy called RESOLFT enables comparatively fast and continuous imaging of sensitive, nanosized features in living brain tissue. Using low-intensity illumination to switch photochromic fluorescent proteins reversibly between a fluorescent and a nonfluorescent state, we increased the resolution more than 3-fold over that of confocal microscopy in all dimensions. Dendritic spines located 10–50 μm deep inside living organotypic hippocampal brain slices were recorded for hours without signs of degradation. Using a fast-switching protein increased the imaging speed 50-fold over reported RESOLFT schemes, which in turn enabled the recording of spontaneous and stimulated changes of dendritic actin filaments and spine morphology occurring on time scales from seconds to hours.

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Language(s): eng - English
 Dates: 2012-09-20
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1016/j.neuron.2012.07.028
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Title: Neuron
Source Genre: Journal
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Pages: - Volume / Issue: 75 (6) Sequence Number: - Start / End Page: 992 - 1000 Identifier: -