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Flattened cochlear preparations were placed in a recording chamber mounted on an upright microscope (Axioskop2 FX plus, Carl Zeiss, Germany).
The isolated neural preparations were placed in an orientation so that the rise of the platform corresponded to a head down tilt in the intact animal.
Aortic preparations were placed in chambers, kept at 37°C and bubbled continuously with 95% O2 – 5% CO2 in Krebs Bicarbonate Buffer (KBF) (pH 7.4).
Aliquots of the sperm preparations were placed on a Makler chamber at 38°C to subjectively assess their progressive motility under a phase-contrast microscope at 200 × magnification.
The brain was covered with 50 μL of the dye solution and preparations were placed inside a box with a wet tissue for 1.5 h.
The preparations were placed in a custom pacing and imaging chamber and voltage mapping of spontaneous cardiac electrical activity was performed using this chamber and a CCD (CardioCCD-SMQ, RedShirtImaging, Decatur, GA) mounted on a Nikon TE2000 inverted microscope.
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7.5 µL of sample solution (0.1 w/v%, diluted from 1 w/v% immediately before grid preparation) were placed on 300-mesh copper grids with lacey carbon support, blotted, and plunge frozen into liquid ethane.
Once dissection was complete, the preparation was placed in a supine position in a specially designed plexiglass chamber and secured with ear bars.
In brief, 50 100 μL PureCeption-washed sperm (see sperm preparation) was placed onto a cleaned uncoated 12-mm circle glass coverslip (Fisher 12-545-81) and allowed to bind for 10 20 min.
This rendered the threshold to sound well above 70 dB SPL, and thus above the background noise in the sound reduced chamber, where the preparation was placed.
The insect preparation was placed one cm apart from the tip of an omni-directional hot-wire anemometer (Ahlborn, Almemo FVA 605 TA5O), which was connected to a calibrated data logger (Ahlborn, Almemo 2690) sampling the actual wind velocity every 20 ms. The preparation was positioned in the middle of the floor of the flight arena.
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