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Kanamycin resistant bacteria were enumerated using digital images of the plates, which were acquired with a Q-Counter (Spiral Biotech, Norwood MA)) after 28 and 48 h of incubation.
The viable counts of nitrifying bacteria were enumerated using the agar medium recommended by Aaronson [ 17] after incubation at 22°C for 10 days.
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To examine cells in varying physiological states, bacteria were enumerated based on the Live/Dead BacLight kit, DAPI (4′,6-diamidino-2-phenylindole) staining, fluorescent in situ hybridization (FISH), and colony forming units (CFU).
At the indicated times post infection, host cells were lysed and bacteria were enumerated for colony forming units.
After washing for 3×, the extracellular bacteria were killed by adding gentamicin (100 μg per mL in PBS for 60 min. Finally, amoebae and bacteria were enumerated as described above. The bacterial colony forming units invading A. castellanii were calculated as follows: (number of bacterial c.f.u./number of amoebae) × 100 = bacterial invasion of A. castellanii.
Agar plates were aerobically incubated (37 °C) for 24 h, following which the bacteria was enumerated and the appropriate inoculation concentrations could be prepared by dilution of the culture in PBS.
Bacteria were stained using immunofluorescence.
The growing evidence supported the notion that the bacteria were being used in a coordinated way.
In this case, no real bacteria were used, but Shendruk said that E. coli bacteria have been commonly used in experimental works.
Samples without bacteria were used as controls.
In medicine, genetically modified bacteria are used to produce insulin.
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