Protocols   Select Another Resource Report Type

Authors: Lixin Wang, Collin Challis, Honghui Liang, Songlin Li, Charless Fowlkes, Aidan Sullivan, Kumar SR, Yvette Taché
Group: SPARC
Summary: Using a multicolor adeno-associate virus system to label the colonic enteric nervous system for digital tracing of individual neurons and nerve fibers in microcircuits in three-dimensions (3D). The methods include viral vectors retro-orbital injection in mice, preparation of colon tissues, microscopy and 3D digital tracing.

Proper citation: Lixin Wang, Collin Challis, Honghui Liang, Songlin Li, Charless Fowlkes, Aidan Sullivan, Kumar SR, Yvette Taché 2020. Multicolor adeno-associate virus labeling and 3D digital tracing of enteric plexus in mouse proximal colon. protocols.io https://dx.doi.org/10.17504/protocols.io.bqavmse6 Copy   

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Authors: Latoya Allen, Marissa Ciesla, Yasin Seven, Elisa Gonzalez-Rothi, Gordon Pool
Group: SPARC
Summary: This protocol describes the procedure for retrogradely labeling bilateral phrenic motor neuron pools in the cervical spinal cord using intrapleural injections of cholera toxin B fragment

Proper citation: Latoya Allen, Marissa Ciesla, Yasin Seven, Elisa Gonzalez-Rothi, Gordon Pool 2019. SPARC Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons Using Intrapleural Injections of Cholera Toxin B Fragment. protocols.io https://dx.doi.org/10.17504/protocols.io.2kpgcvn Copy   

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Authors: Charles Horn, Derek M. Miller, Stephanie Fulton, Bill J. Yates, Lee E. Fisher, Ameya C. Nanivadekar
Group: SPARC
Summary: This protocol is used to surgically implant gastrointestinal planar electrodes and vagus nerve cuff electrodes for chronic electrophysiological recording and stimulaiton in the ferret. Electrode lead wiring is tunneled subcutaneously to a connector embedded in a headcap affixed to the cranium.Funding: This protocol was developed with funding from the NIH Common Fund’s Stimulating Peripheral Activity to Relieve Conditions (SPARC) program (Award U18TR002205). To learn more about the SPARC program, visit https://sparc.science.

Proper citation: Charles Horn, Derek M. Miller, Stephanie Fulton, Bill J. Yates, Lee E. Fisher, Ameya C. Nanivadekar 2019. SPARC - Chronic implantation of gastrointestinal and vagus nerve electrodes in the ferret. protocols.io https://dx.doi.org/10.17504/protocols.io.6crhav6 Copy   

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Authors: Lauren Segers, Kendall Morris, Donald Bolser
Group: SPARC

Proper citation: Lauren Segers, Kendall Morris, Donald Bolser 2019. Morris USF Lab protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.58vg9w6 Copy   

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Authors: Pradeep Rajendran, John Tompkins, Kalyanam Shivkumar
Group: SPARC

Proper citation: Pradeep Rajendran, John Tompkins, Kalyanam Shivkumar 2019. Pig- Heart Neuonal and Fiber Immunocytochemistry. protocols.io https://dx.doi.org/10.17504/protocols.io.2n9gdh6 Copy   

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Authors: Jeffrey Ardell
Group: SPARC
Summary: Yorshire pigs are anthesized for ICN/ neuromodulation recodings. Montior hemodynics A1mid-sternotomy

Proper citation: Jeffrey Ardell 2019. Pig ICN recording. protocols.io https://dx.doi.org/10.17504/protocols.io.2jugcnw Copy   

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Authors: Janet Keast, Peregrine Osborne
Group: SPARC
Summary: This protocol describes confocal microscopy and image analysis procedures for characterizing neuronal cell bodies and their associated synaptic boutons in thick (50 µm) cryosections. The protocol has been applied to rat pelvic ganglia, where neuronal cell bodies have been identified using immunohistochemical markers of specific neuron populations and/or fluorescent retrograde tracer.

Proper citation: Janet Keast, Peregrine Osborne 2020. Confocal microscopy and characterization of synaptic boutons associated with ganglion neurons. protocols.io https://dx.doi.org/10.17504/protocols.io.bakdics6 Copy   

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Authors: James Fallon, Sophie Payne
Group: SPARC
Summary: Recording electrically-evoked compound action potentials (ECAPs) is performed to confirm that the electrode array is appropriately placed along the nerve. Recording ECAPS also allows confirmation that electrical stimulation is above neural threshold i.e. that electrical stimulation activates neural activity. The procedure is performed under anesthesia (immediately post-surgery) or in awake animals, and the procedure should incorporate all local requirements for standards of animal experimentation.

Proper citation: James Fallon, Sophie Payne 2020. Electrophysiological recording of electrically-evoked compound action potentials. protocols.io https://dx.doi.org/10.17504/protocols.io.bfwyjpfw Copy   

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Authors: Maci Heal, Susan Tappan
Group: SPARC
Summary: Convert binary mask tracings from Nikon's NIS Elements software into MBF XML format.

Proper citation: Maci Heal, Susan Tappan 2020. SPARC Segmentation Conversion: Binary mask tracings to MBF XML. protocols.io https://dx.doi.org/10.17504/protocols.io.bchjit4n Copy   

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Authors: Elizabeth Smith
Group: SPARC
Summary: Immunohistochemistry protocol used for staining with fluorescent secondary antibodies to highlight specific tissue structures and amplify specific antibody signals.

Proper citation: Elizabeth Smith 2019. IHC-Amplified Fluorescent Frozen Sections. protocols.io https://dx.doi.org/10.17504/protocols.io.8rmhv46 Copy   

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Authors: Muriel Larauche, Yushan Wang, Po-Min Wang, James Dunn, Wentai Liu, Mulugeta Million
Group: SPARC
Summary: This protocol describes a process for the measurement of acute electrical stimulation-induced effects on colonic motility in anesthetized young adult Yucatan minipigs. Signals recorded from manometry probes inserted into the proximal, transverse and distal colonic regions were used to measure the effect of stimulation by serosally placed electrodes opposed to the colonic tissue at multiple sites in an acute anesthetized preparation. The effect of stimulation was quantified as motility index assessments before, during and after stimulation, and the data was used to create a functional map of colonic motor response to localized colon stimulation.

Proper citation: Muriel Larauche, Yushan Wang, Po-Min Wang, James Dunn, Wentai Liu, Mulugeta Million 2020. Tache_Mulugeta_OT2OD024899_Colon tissue electrical stimulation and colonic motility measurements. protocols.io https://dx.doi.org/10.17504/protocols.io.3rmgm46 Copy   

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Authors: Nicole A. Pelot, J. Ashley Ezzell, Gabriel B. Goldhagen, Jake E. Cariello, Kara A. Clissold, Warren M. Grill
Group: SPARC
Summary: The protocol describes immunohistochemistry with anti-claudin-1, imaging, image segmentation, and image analysis methods to quantify human vagus nerve morphology.

Proper citation: Nicole A. Pelot, J. Ashley Ezzell, Gabriel B. Goldhagen, Jake E. Cariello, Kara A. Clissold, Warren M. Grill 2020. SPARC_Duke_PelotGrill_OT2-OD025340_HumanVagusNerve_Claudin1IHC_Morphology. protocols.io https://dx.doi.org/10.17504/protocols.io.bh4dj8s6 Copy   

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Authors: Elizabeth Butterworth, Wesley Dickerson, Vindhya Vijay, Kristina Weitzel, Julia Cooper, Eric W. Atkinson, Jason E. Coleman, Kevin Otto, Martha Campbell Thompson
Group: Human Cell Atlas Method Development Community, Optical Clearing of Tissue, SPARC
Summary: Using traditional histological methods, researchers are hampered in their ability to image whole tissues or organs in large-scale 3D. Histological sections are generally limited to 500 μm using traditional methods. In addition, light scatters from macromolecules within tissues, particularly lipids, prevents imaging to a depth >150 μm with most confocal microscopes. To reduce light scatter and to allow for deep tissue imaging using simple confocal microscopy, various optical clearing methods have been developed that are relevant for rodent and human tissue samples fixed by immersion. Several methods are related and use protein crosslinking with acrylamide and tissue clearing with sodium dodecyl sulfate (SDS). Other optical clearing techniques used various solvents though each modification had various advantages and disadvantages. Here, an optimized passive optical clearing method is described for studies of the human pancreas innervation and specifically for interrogation of the innervation of human islets.

Proper citation: Elizabeth Butterworth, Wesley Dickerson, Vindhya Vijay, Kristina Weitzel, Julia Cooper, Eric W. Atkinson, Jason E. Coleman, Kevin Otto, Martha Campbell Thompson 2020. Human Pancreas PACT Optical Clearing and High Resolution 3D Microscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.9gbh3sn Copy   

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Authors: Nathan Lee
Group: SPARC

Proper citation: Nathan Lee 2020. Abdominal Emitter Implantation. protocols.io https://dx.doi.org/10.17504/protocols.io.bpzbmp2n Copy   

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Authors: Lauren Segers, Kendall Morris, Sarah Nuding
Group: SPARC

Proper citation: Lauren Segers, Kendall Morris, Sarah Nuding 2019. Morris USF Lab protocol DEMO. protocols.io https://dx.doi.org/10.17504/protocols.io.5gbg3sn Copy   

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Authors: J. Ashley Ezzell, Nicole A. Pelot, Kara A. Clissold, Warren M. Grill
Group: SPARC
Summary: Protocol for collection, histological processing, and imaging of rat vagus nerves.

Proper citation: J. Ashley Ezzell, Nicole A. Pelot, Kara A. Clissold, Warren M. Grill 2019. SPARC_Duke_Grill_OT2-OD025340_RatVagusNerveCollectionHistologyMicroscopy. protocols.io https://dx.doi.org/10.17504/protocols.io.ww3ffgn Copy   

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Authors: Elizabeth Smith, Jacqui Potter
Group: SPARC
Summary: ImageJ Software can be used to find the percentage of nerve density in regions of interest. These regions of interest are imaged, and the nerve percentage is found by adjusting the image's histogram threshold.

Proper citation: Elizabeth Smith, Jacqui Potter 2020. Image J Nerve Density Percentage Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.beh4jb8w Copy   

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Authors: Janet Keast, Peregrine Osborne
Group: SPARC
Summary: This protocol is used to visualise sensory and autonomic neurons innervating the bladder body (dome), bladder trigone or proximal urethra in an experimental adult male or female rat. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.

Proper citation: Janet Keast, Peregrine Osborne 2020. Use of tracer dyes to label neural projections to lower urinary tract organs. protocols.io https://dx.doi.org/10.17504/protocols.io.w2xfgfn Copy   

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Authors: Deborah Jaffey, Logan Chesney, Terry Powley
Group: SPARC
Summary: This protocol describes how to prepare a rat stomach for micro-CT imaging of the vasculature. A lead-containing silicone-based curable material is injected into the heart at perfusion and flows through the vasculature and cures in place. Subsequent microCT imaging then provides a 3D description of the stomach vasculature.

Proper citation: Deborah Jaffey, Logan Chesney, Terry Powley 2020. Micro-CT imaging of rat stomach vasculature. protocols.io https://dx.doi.org/10.17504/protocols.io.bafnibme Copy   

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Authors: J. Ashley Ezzell, Nicole A. Pelot, Kara A. Clissold, Warren M. Grill
Group: SPARC
Summary: The protocol describes immunohistochemistry with anti-choline acetyltransferase, as it has been applied to cervical and abdominal vagus nerve samples from rats, pigs, and humans.

Proper citation: J. Ashley Ezzell, Nicole A. Pelot, Kara A. Clissold, Warren M. Grill 2020. SPARC_Duke_Grill_OT2-OD025340_VagusNerve_IHC_ChAT. protocols.io https://dx.doi.org/10.17504/protocols.io.bi9tkh6n Copy   

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