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Mice were anesthetized with 0.5% sodium pentobarbital, and livers were digested with a 2-step perfusion method using liver perfusion and digestion media (Invitrogen) delivered through the portal vein.
Briefly, the livers were digested with 4% KOH overnight, and the number of eggs was determined by microscopic examination.
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After de-waxing and post-fixation, paraffin embedded sections of mice liver were digested with 10 µg/ml Proteinase K in 0.1 M Tris pH 7.5 for 10 min at 37°C.
DNA, isolated from the upper gastrointestinal tract and liver, was digested to 3'-nucleotides.
Except for 3-week old rats, where the whole liver was used per rat, one lobe of the liver was digested for each of the older rats.
Liver was digested in 2 mg/mL collagenase D (Sigma-Aldrich) at 37°C for 30 minutes and then passed through a 100-μm filter.
Lungs, livers and brains were digested with digestion buffer, 1 mg/ml collagenase (Wako Pure Industries, Ltd, Osaka, Japan), 1 mg/ml dispase (Gibco) and DNase (Sigma-Aldrich) in PBS at 37° C for 30 minutes.
Liver samples were digested in 1× PBS supplemented with DNase I and type I/II collagenases for 15 min at 32°C.
Five μm paraffin-embedded liver sections were digested with trypsinase for 30 min at 37°C, followed by the antigen retrieval via pressure cooking for 2 min in 0.01 mol/L citrate buffer (PH 6.0).
For liver cell and lung cell preparation, livers and lungs were digested with collagenase D and DNase I, passed through a sterile 40 µm pore size nylon cell strainer (BD Biosciences) and the cell suspensions were treated with RBC lysis buffer (eBioscience).
Dissected liver and CNS tissues were digested 15 mins at 37°C in trypsin-EDTA, heart 30 mins at 37°C in trypsin-EDTA followed by 30 mins in collagenase.
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