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For in vivo B cell analysis, lymphoid organs were cut into small pieces, treated with collagenase D (Boehringer, Mannheim, Germany) for 30 min at 37°C, gently meshed and washed with PBS containing 50 µg/mL Dnase I (Roche, Mannheim, Germany) and 2 mM EDTA.
For leaf pieces treated with a 30 mM NaOH solution, the mean time for cell death was 33.72 ± 5.44 min.
The cells of leaf pieces treated with 3 mM HCl solution died 4.02 ± 1.02 h following treatment.
For leaf pieces treated with 2 M NaCl, cell colouration changed gradually and eventually became green as cell death progressed over a 4.25 ± 0.38 h timeframe.
Leaf pieces treated with 400 mM NaCl, cell colouration appeared grey until turning greener at the moment of cell death after 5.32 ± 0.63 h.
For leaf pieces treated with a 1 M NaOH solution, the mean time for induced cell death was 49 ± 5.3 s.
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The hide piece treated with 50 mM Tris buffer (pH 8.0) instead of protease served as negative control.
For preparing viable glioma cells, freshly obtained specimen were cut finely into small pieces, treated in IMDM with 0.5 mg/ml collagenase (Sigma) and 25 µg/ml DNAse (Sigma) at 37°C for 40 minutes.
Chicken meat pieces were treated with maltodextrin (0.036 g/g meat) or maltodextrin/2IB3MP (0.075 g/g meat).
About 10 ml mesoglea jellyfish from each population (BsA, JsA, WsA) was cut into pieces and treated with 1 mg/ml collagenase (ICN, cat N 150705) solution in sea water at 37 °C for 30 min.
Tumor tissue (∼200 300 mg) was washed twice with ice-cold Ca2+-, Mg2+-free phosphate-buffered saline (PBS, pH 7.4), minced to small pieces and treated with solution of 0.125% Trypsin/0.015% EDTA in PBS containing 250 µg/ml DNAse I.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com