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Double-sided desheathing of a leech ganglion.
DOI: 10.21769/BioProtoc.v460 Published: Mar 5, 2018 Views: 3980
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1. Position a disk of transparent PDMS in the center of the container. 2. Transfer the ganglion into a microdissection container filled with cold saline. 3. Place the ganglion over the disk of transparent PDMS. 4. Use short pieces of tungsten wire to pin down sinus (the ganglion’s ventral side is up). 5. Stretch the connectives to apply tension to the ganglion’s sheath. 6. Zoom in on the target ganglion. 7. Remove the ventral sheath from the glial packet in the following order: the posterior, the central, and the lateral glial packets. 8. Check that the cells are not displaced or damaged. 9. Unpin the ganglion, flip the dorsal side up, then pin it down. 10. Zoom in on the ganglion. 11. Remove the dorsal sheath from the lateral glial packets (posterior to anterior). 12. Check that the cells are not displaced or damaged. 13. Unpin the ganglion, flip the ventral side up, then pin it down. 14. Check that the cells are not displaced or damaged.
Extracted from protocol
Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia
Tomina, Y. and Wagenaar, D. A. (2018). Bio-protocol 8(5): e2751.
DOI: 10.21769/BioProtoc.2751.
1. Position a disk of transparent PDMS in the center of the container. 2. Transfer the ganglion into a microdissection container filled with cold saline. 3. Place the ganglion over the disk of transparent PDMS. 4. Use short pieces of tungsten wire to pin down sinus (the ganglion’s ventral side is up). 5. Stretch the connectives to apply tension to the ganglion’s sheath. 6. Zoom in on the target ganglion. 7. Remove the ventral sheath from the glial packet in the following order: the posterior, the central, and the lateral glial packets. 8. Check that the cells are not displaced or damaged. 9. Unpin the ganglion, flip the dorsal side up, then pin it down. 10. Zoom in on the ganglion. 11. Remove the dorsal sheath from the lateral glial packets (posterior to anterior). 12. Check that the cells are not displaced or damaged. 13. Unpin the ganglion, flip the ventral side up, then pin it down. 14. Check that the cells are not displaced or damaged.
Extracted from protocol
Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia
Tomina, Y. and Wagenaar, D. A. (2018). Bio-protocol 8(5): e2751.
DOI: 10.21769/BioProtoc.2751.
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Keywords
Voltage-sensitive dye imaging, Whole-brain recording, VoltageFluor, Microdissection, Dye-loading process
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