Prior to dissection, animals were placed on ice for at least 30 min. Dissection of the crab stomatogastric nervous system (STNS) were performed in two parts, the gross and the fine dissection. Breifly, in the gross dissection the stomach was dissected from the animal. Subsequently in the fine dissection we removed the intact stomatogastric nervous system (STNS) from the stomach under a dissection microscope using fine micro-dissection tools. The STNS includes the commissural ganglia, esophageal ganglion, and STG with connecting motor nerves (Gutierrez and Grashow, 2009 (link)). The STNS was pinned in a Sylgard-coated (Dow Corning) dish and continuously superfused with 11 °C saline.
Sylgard coated
Manufactured by Dow
Sylgard-coated lab equipment is a line of laboratory equipment with a silicone-based coating. The coating helps to protect the equipment from chemical exposure and corrosion. The core function of Sylgard-coated equipment is to provide a durable and chemically-resistant surface for laboratory applications.
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Lab products found in correlation
7 protocols using sylgard coated
1
Crab Stomatogastric Nervous System Dissection
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Dissection of Jonah Crab Central Ganglia
3
Intact Olfactory Network Brain Preparation
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The brain preparation leaving the entire olfactory network intact has been described previously (Demmer and Kloppenburg, 2009 (link); Husch et al., 2009a (link); Kloppenburg et al., 1999 (link)). Animals were anesthetized by CO2, placed in a custom-built holder, and the head was immobilized with tape (tesa ExtraPower Gewebeband, Tesa, Hamburg, Germany). The head capsule was opened by cutting a window between the two compound eyes and the antennae’s bases. The brain with its antennal nerves and attached antennae was dissected in extracellular saline (see below) and pinned in a Sylgard-coated (Dow Corning Corp., Midland, MI) recording chamber. To get access to the recording site, we desheathed parts of the AL using fine forceps, and preparations were enzymatically treated with a combination of papain (0.3 mg·ml–1, P4762, Sigma) and L-cysteine (1 mg·ml–1, 30090, Fluka) dissolved in extracellular saline (~3 min, room temperature [RT], ~24°C). For electrophysiological recordings, the somata of the AL neurons were visualized with a fixed stage upright microscope (AxioExaminer, Carl Zeiss, Jena, Germany) using a 20× water-immersion objective (20× W Apochromat, NA = 1) with a 4× magnification changer, and infrared differential interference contrast optics (Dodt and Zieglgänsberger, 1994 (link)).
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Dissection of Jonah Crab STNS
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Adult male Jonah Crabs, Cancer borealis, (N = 43) were obtained from Commercial Lobster (Boston, MA) and maintained in artificial seawater at 10°C–12°C in a 12 hr light/dark cycle. On average, animals were acclimated at this temperature for 1 week before use. Prior to dissection, animals were placed on ice for at least 30 min. Dissections were performed as previously described (Gutierrez and Grashow, 2009 (link)). In short, the stomach was dissected from the animal and the intact stomatogastric nervous system (STNS) was removed from the stomach including the commissural ganglia, esophageal ganglion and stomatogastric ganglion (STG) with connecting motor nerves. The STNS was pinned in a Sylgard-coated (Dow Corning) dish and continuously superfused with saline. Saline was composed of 440 mM NaCl, 11 mM KCl, 26 mM MgCl2, 13 mM CaCl2, 11 mM Trizma base, 5 mM maleic acid, pH 7.4–7.5 at 23°C (~7.7–7.8 pH at 11°C).
5
Crustacean Nervous System Dissection
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Adult male crabs (Cancer borealis) were acquired from local distributors and maintained in aquaria filled with chilled (12–13°C) artificial sea water until use. Crabs were anesthetized before dissection by placing them in ice for at least 20 min. The stomatogastric nervous system including the STG, esophageal ganglion, the pair of commissural ganglia, and the motor nerves were dissected from the stomach and pinned to a saline filled, Sylgard-coated (Dow Corning) Petri dish (schematic in Fig. 1A ). The STG was desheathed, exposing the somata of the neurons for intracellular impalement. Preparations were superfused with chilled (10–13°C) physiological saline containing: 11 mm KCl, 440 mm NaCl, 13 mm CaCl2 · 2H2O, 26 mm MgCl2 · 6H2O, 11.2 mm Tris base, and 5.1 mm maleic acid with a pH of 7.4.
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Dissection and Isolation of Crab Cardiac Ganglion
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Male Jonah crabs, C. borealis, were obtained from Commercial Lobster (N = 60 crabs; Boston, MA) and housed in artificial seawater tanks at 10–12˚C. On average, animals were acclimated to this temperature for one week before use. Prior to dissection, animals were placed on ice for at least 30 min. Dissections were performed as previously described (Cruz‐Bermúdez & Marder, 2007 (link)). In short, the heart was dissected from the animal and the intact CG was removed, preserving the connections between the small cells and the large cells. The CG was pinned in a Sylgard‐coated (Dow Corning) dish in physiological saline.
7
Isolation and Dissection of Crab Stomatogastric Nervous System
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Adult male crabs (Cancer borealis) were acquired from local distributors and maintained in aquaria filled with chilled (12-13°C) artificial sea water until use. Crabs were anesthetized before dissection by placing them in ice for at least 20 minutes. The stomatogastric nervous system including the stomatogastric ganglion (STG), esophageal ganglion, the pair of commissural ganglia, and the motor nerves were dissected from the stomach and pinned to a saline filled, Sylgard-coated (Dow Corning) Petri dish (schematic in Figure 1A). The STG was desheathed, exposing the somata of the neurons for intracellular impalement. Preparations were superfused with chilled (10-13°C) physiological saline containing: 11 mM KCl, 440 mM NaCl, 13 mM CaCl 2 • 2H 2 O, 26 mM MgCl 2 • 6H 2 O, 11.2 mM Tris base, 5.1 mM maleic acid with a pH of 7.4.
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