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After reaching confluence, these cells were trypsinized, centrifuged, suspended in media, and counted.
The exact amount of inoculated cells or spores in the different products was checked in preliminary work where inoculation suspensions were plated on specific media and counted afterward.
Cell suspensions were washed with complete media and counted.
The cells were then trypsinized, re-suspended in the media and counted.
For proliferation experiments, 104 cells were plated in growth media and counted via a hemocytometer 72, 144, and 196 hours after plating.
Furthermore, promastigotes were treated with the same inhibitor at different days of culture, washed, re-seeded in new media and counted after 6 days of culture.
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Depending on the nature and energy of the reaction products (from a few-keV Auger electrons up to 8 MeV gamma rays) that the detector aims at isolating as an indirect neutron signature, a variety of sensor media and counting methods have been introduced during the last decades.
The bacteriological method, which is generally used for assessing the multiplicity of mycobacterial infection in animal organs and tissues, involves inoculation of their homogenates on special agar media and counting colony-forming units.
Then, the cells were detached by gentle agitation, washed off with culture media, collected and counted in Neubauer hemocytometric chamber in presence of trypan blue dye to distinguish between live and dead cells in suspension.
The cells that migrated into the lower chamber media were collected and counted by the Bio-Rad (Hercules, CA, USA) TC10 automatic cell counter.
To count, cells were washed with Hanks, trypsinized, centrifuged, and resuspended in 5 ml of media and then counted manually using Haemocytometer every day for 7 days.
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