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The excised brain was washed in ice-cold saline and subsequently, one hemisphere was homogenized in 12 mL ice-cold saline and centrifuged for 5 min at 4,000 rpm.
After incubation, the brain was washed three times with 5 min intervals in PBS.
Subsequently, the brain was washed in 0.9% normal saline to remove the blood from the surface, put into a 25 mL glass beaker filled with saline, and maintained at 37 degrees Celsius.
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The sections from the entire brain were washed in PBS-0.2% Triton and treated for 30 min with H2O2 (0.3%).
For brain sections: sections from the entire brain were washed in PBS-0.2% Triton and blocked with goat serum (1 100 in PBS, Vector) for 60 min.
On the other hand, when a fraction of the glucose and lactate taken up from blood into brain is washed out of brain via perivascular fluid flow before it is used by brain cells, then uptake and metabolic rates calculated by the Fick principle (i.e. CBF[A-V]) will be overestimated.
Then, the brains were washed with PBS.
After primary antibody incubation, brains were washed in 0.5% PBT for 6 × 30 min at RT.
Next, the brains were washed with PBS, mounted on Superfrost Plus glass slides, and mounted with Mowiol.
But it does more than that: it gives a shape to a process that we then might easily take for granted: can brains be "washed"?
Brains were washed in 0.5% PBT for 6 × 30 min at RT followed by overnight incubation in Vectashield® (Vector Laboratories) before confocal scanning.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com