CIL: 54356
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- Collection
- Cite This Work
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Petersen, Mark. A.; Tognatta, Reshmi; Meyer-Franke, Anke; Bushong, Eric. A.; Mendiola, Andrew. S.; Yan, Zhaoqi; Muthusamy, Abinaya; Merlini, Mario; Meza-Acevedo, Rosa; Cabriga, Belinda; Ryu, Jae Kyu; Lassmann, Hans; Ellisman, Mark. H.; Akassoglou, Katerina (2022). CIL: 54356. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0BK1C3P
- Description
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Following in vivo 2-photon imaging of an area of spinal cord with chronic experimental autoimmune encephalomyelitis, a correlated block-face SEM volume was collected of an area near blood vessels displaying clustering of glial cells (either oligodendrocytes, microglia, or both) or control blood vessels lacking clustering.
- Date Issued
- 2022
- Researchers
- Methods
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The animal was perfused with Ringer's solution followed by 0.5% glutaraldehyde / 2% PFA in cacodylate. The region of spinal cord under the imaging window was cut from the perfused cord and post-fixed for 2 hours in cold 0.5% glutaraldehyde / 2% PFA in cacodylate. The specimen was then post-fixed overnight in cold 4% PFA in cacodylate. The dorsal aspect of the cord was cut into 150 µm thick horizontal vibratome sections. The sections were post-fixed overnight in cold 2% glutaraldehyde in cacodylate overnight. The tissue was stained with 2% osmium tetroxide (Ted Pella) in 0.15M cacodylate, 0.5% aq. thiocarbohydrazide (Electron Microscopy Sciences), 2% aq. osmium tetroxide, 2% aq. uranyl acetate (Ted Pella), and lead aspartate, with thorough washing with water between each staining solution. The sections were then dehydrated through ethanol and acetone and then infiltrated with Durcupan ACM (Millipore Sigma). The sections flat-embedded between glass slides coated with mold-release compound (Electron Microscopy Sciences, Hatfield PA) and were cured at 60 °C for 72 hours. Specimens were imaged on a Zeiss Gemini 300 VP SEM equipped with a focal charge compensation system and a Gatan 2XP 3View system. Volumes were collected at 2.5 kV with 1 µsec dwell time, 10 nm pixels, 50 nm step size, and focal gas injection with nitrogen gas turned on. The scope was run in analytic mode and high current mode. The resulting stacks of images were aligned using a custom Python script using IMOD programs.
- Technical Details
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Imaging mode: serial block face SEM (SBFSEM)
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- Scientific Name
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- Language
- No linguistic content; Not applicable
- Identifier
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Samplenumber: 54356
- Related Resource
- Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL54356
Source data
- License
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Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International Public License
- Rights Holder
- UC Regents
- Copyright
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Under copyright (US)
Use: This work is available from the UC San Diego Library. This digital copy of the work is intended to support research, teaching, and private study.
Constraint(s) on Use: This work is protected by the U.S. Copyright Law (Title 17, U.S.C.). Use of this work beyond that allowed by "fair use" or any license applied to this work requires written permission of the copyright holder(s). Responsibility for obtaining permissions and any use and distribution of this work rests exclusively with the user and not the UC San Diego Library. Inquiries can be made to the UC San Diego Library program having custody of the work.
- Digital Object Made Available By
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Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp)
- Last Modified
2022-11-18