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A steep thermal gradient on a thin agar plate was established as described previously [13].
We created a thermotaxis setup as described previously using a steep thermal gradient on a thin agar plate [20].
Specimens for SEM were prepared by excising thin agar pieces from an actively growing culture and viewed on a Philips XL30 SEM TMP (FEI Company, Hillsboro, OR).
For these reasons, we designed a novel hybrid micro fluidic device that combines a thin agar pad and a custom flow chamber.
Until now, most time lapse microscopy experiments of bacterial processes ie growth, cell cycle and motility studies have used thin agar pads to provide a substratum that supports growth, starvation or surface motility [5], [12], [15], [16].
Because of these problems and the fact that the agar pad is almost a "universal" substratum in microbiology, we aimed to develop a novel hydrid micro fluidic device that combines a thin agar pad and a custom flow chamber.
Similar(48)
1 µL of a 1×107 cfu/mL exponentially growing E. coli suspension cultures was spotted under a thin fresh agar as described above.
Most time lapse microscopy experiments studying bacterial processes ie growth, progression through the cell cycle and motility have been performed on thin nutrient agar pads.
Thin layer agar (TLA) culture, which is also being explored as an alternative to automated liquid culture in high burden countries, is similar to MODS in that cultures are examined for microcolony growth by microscopic analysis, but is performed on solid medium [9].
These isolates had previously been tested for susceptibility to rifampin and isoniazid using a screening test in thin layer agar Middlebrook 7H11 (TLA 7H11) (Becton Dickinson) [ 24] and confirmed as MDR-TB using the Multiple Proportion Method in agar [ 25].
Cultures were prepared as described above and 5 μl cell suspensions were spot-inoculated on thin R2A agar plates (1 1.5 mm thick; 10 ml in petridish with 9 cm diameter).
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