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In this work, a subset of the publicly available NASA Structure and Liftoff in Combustion Experiments (SLICE) microgravity and normal gravity nitrogen-diluted methane flames has been considered, and a method to extract quantitative CH* concentration information from the SLICE raw data is demonstrated.
In some experiments, slice cultures were pretreated Suc-LLVY-AMC (80 µM) for 30 min before UCB added and calpain activity was assayed from 0 to 60 min after UCB application.
In a subset of experiments, slice cultures (n = 5) were mounted near the top of an aCSF column in a 500 mL beaker containing immersed US transducers, which were affixed to the bottom beakers to provide a 45 mm standoff distance.
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In some experiments slices were pre-washed in ACSF before the measurement of Aβ1 42 release into ACSF.
For experiments, slices were then transferred into a recording chamber where they were continuously perfused (2 3 mL/min) with oxygenated ACSF warmed to 32°C.
In the first set of experiments, slices of fluorescent SVZ, obtained from YFP transgenic mice were cultivated in contact with wild-type cortex (Figure 4A).
In physiology experiments, slices are typically cut from the brain and allowed to recover from the slicing process for at least 30 minutes, often several hours, before further use.
For AHP experiments, slices were then transferred to a heated (32°C) interface-type chamber, maintained in oxygenated (95% O2, 5% CO2) normal-calcium ACSF containing 2mM Cand2mMnd 2mM MgCl2 for least 2 h prior to recording.
For patch-clamp experiments, slices were transferred to the recording chamber perfused with ACSF at a rate of 1 ml/min using a perfusion pump 2232 Microperpex S (LK.B, Upsalla, Sweden).
In these experiments, slices were prepared 7 8 days later.
In lowered [Mg2+] experiments slices were incubated in ACSF containing approximately 100 μM.
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