Worms (1- to 2-day-old) were collected from a 6 cm NGM plate and washed 3 times. Serotonin creatinine sulfate monohydrate (10 mg/mL; Sigma, H7752) was added to the worm pellet and incubated for 10 min at 20 to 25°C. Worms were loaded onto NGM plates, and a cover glass was placed. Tooth movement was observed at 400× magnification and recorded using a CCD camera (Leica, MC170) at 30 fps for 15 s.
H7752
Manufactured by Merck Group
Sourced in Sao Tome and Principe
The H7752 is a laboratory equipment product manufactured by Merck Group. It is designed to serve as a general-purpose instrument for various scientific applications. The core function of the H7752 is to provide a controlled and consistent environment for experiments or sample processing. Further details on the specific intended use or features of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using h7752
1
Serotonin-Induced Tooth Movement in Worms
2
Quantitative Electropharyngogram Assay of Anthelmintics
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All EPG recordings were made in M9 buffer (Stiernagle, 2006 ), to which drugs, solvents or bacteria were added. Stocks of 5-Hydroxytryptamine creatinine sulfate complex (5HT, Sigma-Aldrich H7752; St. Louis, MO) were prepared in M9 at 40 mM, stored at −20 °C and diluted to desired concentrations in M9. Chip perfusion was initiated within 70 min of preparing a 5HT solution. Ivermectin (IVM; Sigma-Aldrich I8898) stocks (5 mM) were prepared in 100% DMSO and stored at −20 °C. The highest concentrations of DMSO that did not perturb EPG activity in control experiments were 0.2% (Fisher D-136) or 0.5% (Sigma-Aldrich Hybri-Max D2650) (data not shown) so these were the highest concentrations used in working solutions. Levamisole hydrochloride (LEV; Sigma 31,742) stocks (100 mM) were prepared in dH20 and stored at 4 °C until dilution on the day of use. Piperazine hexahydrate (PPZ; Sigma P7003) stocks (1M) were prepared in dH20 and stored at 4 °C until use. The pH of PPZ solutions was adjusted to 7.0 using HEPES buffer in M9 and titrating with 12 N HCl. Working solutions of all anthelmintic drugs were prepared from stocks and used on the same day. In some initial experiments, 0.005% Fast Green (Fisher F-99) was added to solutions to confirm uninterrupted perfusate flow (Lockery et al., 2012 (link)).
3
Serotonin-Induced Egg Laying and Calcium Imaging
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Egg laying in response to exogenous serotonin was performed as described (Banerjee et al., 2017; Kopchock et al., 2021) . Individual staged adult animals were placed in 100 μl of either M9 buffer alone, or M9 containing 18.5 mM serotonin (creatinine sulfate monohydrate salt, Sigma-Aldrich #H7752) or M9 containing 10 µM PMA (Phorbol-12-myristate-13-acetate, Calbiochem #524400) in a 96-well microtiter dish. After 1 hour, the number of released eggs and L1 larvae in each well were counted. Since egg-laying defective animals sometimes release one or two eggs/L1 larvae in response to mechanical stimulation when they are first picked into the well, animals were only recorded as responding if they laid 3 or more progeny. For calcium imaging, NGM plates containing either PMA or ethanol solvent were prepared as described (Reynolds et al., 2005) . Age-matched adult worms from each genotype were placed on separate PMA or control NGM plates at room temperature for 1.5 hour. An agar chunk was then placed between two glass coverslips for Ca 2+ activity recording as described (Ravi et al., 2018b) . The unused plates were kept at 4 °C for future use.
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