Aug 24, 2023
Histology and Retention of Implanted Carbon Fiber Thread Electrodes in Fixed Tissue
- Helen N Schwerdt1,2,
- Tomoko Yoshida3,
- Ann M Graybiel3
- 1Department of Bioengineering, University of Pittsburgh;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD;
- 3McGovern Institute and Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology
Protocol Citation: Helen N Schwerdt, Tomoko Yoshida, Ann M Graybiel 2023. Histology and Retention of Implanted Carbon Fiber Thread Electrodes in Fixed Tissue . protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr4nm3gmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 11, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 80334
Keywords: ASAPCRN
Funders Acknowledgement:
NIH NINDS R00
Grant ID: R00 NS107639
BBRF NARSAD Young Investigator Grant
Grant ID: 28690
Michael J. Fox Foundation for Parkinson’s Research (MJFF) and the Aligning Science Across Parkinson’s (ASAP) initiative
Grant ID: ASAP-020-519
NIH NIMH R01
Grant ID: R01 MH060379
Saks Kavanaugh Foundation
William N. and Bernice E. Bumpus Foundation
Abstract
Methods to cut and retain implanted carbon fiber electrodes in brain tissue are described.
Rats were deeply anesthetized with Euthasol (pentobarbital sodium and phenytoin sodium from Virbac AH Inc.), then transcardially perfused with 0.9% saline solution followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB).
The implanted electrodes were cut just above the skull and below the copper wire bundle component of the device with a rotary saw (Dremel) to retain the flexible Py-CF portion of the device in the fixed brain tissue.
Brains were removed from the skull, post-fixed for 24 hours in the same fixative solution and stored in 25% glycerol with 0.075% sodium azide in 0.1 M PB overnight or until cutting.
To prepare sections, brains were frozen on dry ice then cut in the transverse plane on a freezing microtome with a microtome knife (C. L. Sturkey, Inc. #K185A).
Sections were cut sequentially, with four sections at 25-μm then one section at 100-μm, repeated until the end of the frozen block of brain.
The 100-μm sections were immediately mounted onto glass slides and air dried, whereas the 25-μm sections were stored in 0.1% sodium azide in 0.1 M PB at 4°C until use.