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Different pH regulators divided into acidifying agent, alkaline and buffer salt with the same addition was selected and applied to the RSHH medium, respectively.
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This is likely explained by the property of cloud point buffers to change the conformation and local ion concentration by specific interaction of the buffer salts with the polymer in the shell.
The maximum score for each parameter was 4. The colons were excised, placed in calcium/magnesium-free Hank's buffered salt solution with 1% FBS (Gibco, Grand Island, NY, USA) and cleaned of the contents.
This was followed by perfusion with 0.05% collagenase (C5138; Sigma-Aldrich) in Hanks' buffered salt solution with Ca2+ and Mg2+ (Sigma-Aldrich) for 3 5 min.
After 24 and 48 h of incubation, we washed cells in PBS, and suspended 1 × 10 cells per ml in Hank's buffered salt solution with 50 n M tetramethylrhodamine methyl ester (TMRM) (Invitrogen).
Cells were loaded with 3 µM TMRM (Sigma, T5428) in Hank's buffered salt solution (HBSS with Ca2+, Sigma, H8264) for 15 min at 37°C.
The coverslips were washed 4 times with Hank's buffered salt solution and fixed with freshly made fixative (methanol-glacial acetic acid, 3 1, v/v) for 5 min, followed by air-drying and 15 min Giemsa stain.
Additional file 1: Figure S4a depicts unreacted buffer salts with cubic structures.
Epithelial cells were removed by incubating intestinal tissues in Hank's buffered salt solution (HBSS) supplemented with EDTA and DTT, followed by extensive washing with PBS.
The cells were washed with Hank's buffered salt solution (HBSS, Life Technologies) and live cell imaging was performed using a fluorescent microscope.
Adherent macrophages were washed 3 times with warm Hanks' buffered salt solution (HBSS, Sigma).
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