For volatile anesthetic treatment, coverslips in 12-well plates with a low volume of culture media (500μl to facilitate gas diffusion, were placed in airtight, humidified modular chambers (Billups-Rothenberg, Del Mar, CA, USA) as previously described (Mintz, et al. 2013 (link); Xu, et al. 2018b (link)). The chamber was connected to an agent-specific calibrated vaporizer (SuperaVet, Vaporizer Sales and Services Inc, Rockmart, GA, USA) that delivered 2.4% isoflurane mixed with 5% carbon dioxide / 95% air carrier gas at 12 L/min. Carrier gas alone was used as for controls. Gas composition was measured periodically using a 5250 RGM gas analyzer (Datex-Ohmeda, Madison, WI, USA). In some cases, co-treatment or pre-treatment with pharmacologic compounds was performed. These compounds included: TCS 1205 (10 nM, 100 nM, 1μM, Tocris), TCS 1105 (10 nM, 100 nM, 1μM, Tocris), Zolpidem (10 nM, 100 nM, 1μM, Tocris), Bumetanide (10μM, Tocris), and Chloride Ionophore I (3μM, Millipore). After a 15-min equilibration, the sealed chambers with dissociated cultures were placed in an incubator to maintain temperature at 37°C for 1 hour, followed by 20 min exposure to either vehicle control or a recombinant soluble axon guidance cue, Semaphorin 3A (R & D Systems, Bend, OR) at 100 ng/ml to induce growth cone collapse (Figure 1A , B ).
5250 rgm gas analyzer
Manufactured by GE Healthcare
Sourced in United States
The 5250 RGM gas analyzer is a compact and versatile device designed for the measurement and analysis of various gases. It features advanced sensor technology to provide accurate and reliable gas concentration data. The core function of the 5250 RGM is to analyze and monitor the composition of gases, making it a useful tool in a wide range of industrial and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Bumetanide, by Bio-Techne (1 mentions)
Tcs 1105, by Bio-Techne (1 mentions)
Zolpidem, by Bio-Techne (1 mentions)
Semaphorin 3a, by R&D Systems (1 mentions)
Sevoflurane, by GE Healthcare (1 mentions)
Isoflurane, by GE Healthcare (1 mentions)
2 6 diisopropylphenol, by Merck Group (1 mentions)
Sevoflurane, by Abbvie (1 mentions)
2 protocols using 5250 rgm gas analyzer
1
Volatile Anesthetic Exposure and Growth Cone Collapse
2
In Vitro Anesthesia Exposure Protocol
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Coverslips in 12-well plates were placed in identical air-tight, humidified chambers (Billups-Rothenberg, Del Mar, CA, USA) as previously described [43 (link)]. Isoflurane (Baxter Healthcare Cooperation, Deerfield, IL, USA) or sevoflurane (AbbVie Inc., North Chicago, IL, USA) were delivered using an agent-specific, calibrated inline vaporizer (SuperaVet, Vaporizer Sales and Services Inc., Rockmart, GA, USA), and were diluted in 5% CO2/95% O2 carrier gas. Controls for these experiments received 5% CO2/95% O2 carrier gas only. There was a 15-min equilibration period, which was required to achieve the correct concentration of Isoflurane or sevoflurane as measured by a 5250 RGM gas analyzer (Datex-Ohmeda, Madison, WI, USA). Then the sealed chambers were placed in an incubator to maintain temperature at 37 °C for the duration of anesthesia exposure. Isoflurane/sevoflurane concentration was periodically measured at the end of the experimental period to verify that it was appropriately maintained throughout the exposure.
The propofol exposure was done by adding pure 2,6-diisopropylphenol (Sigma Aldrich, Saint Louis, MO, USA) (1 nM, 2 nM, 4 nM) into experiment wells, and incubated at 37 °C for the duration of anesthesia exposure. The exposure was terminated by removing all the media and by adding a combination of previously-removed media without propofol and fresh media.
The propofol exposure was done by adding pure 2,6-diisopropylphenol (Sigma Aldrich, Saint Louis, MO, USA) (1 nM, 2 nM, 4 nM) into experiment wells, and incubated at 37 °C for the duration of anesthesia exposure. The exposure was terminated by removing all the media and by adding a combination of previously-removed media without propofol and fresh media.
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