Biofilms attached to artificial substrates were detached with a sterile soft toothbrush in 400 mL sterile distilled water, and then treated with probe sonication as follows: ultrasonic time 3 s, interval time 8 s, repetition 75 times, power 600 W (Cai et al. 2013, 2014).
From 30-day old plants, leaves were detached with a sharp razor, and gently inserted pair-wise into 0.5 ml semi-solid water with 0.5% agar in a 0.5-ml reaction tube.
At this time, cylinders were removed from the mice, washed and adherent cells (>90% macrophages) were detached with a cell scraper at 4°C, counted, and characterized by flow cytometry.
For sample preparation, cultures were detached with a scraper in PBS then centrifuged.
Cells were detached with a non-enzyme cell dissociation solution, washed, and resuspended in RPMI-1640 medium.
After 2 h, they were detached with a scraper and RNA was extracted using RNeasy MicroKit (Quiagen, Hilden, Germania) [ 24– 27].
After 30 min, the nonadherent cells were removed, and RGCs were detached with a 0.125% trypsin solution.
The next morning, non-migrated cells were detached with a cotton swab, membranes were removed, fixed in methanol, and stained with Diff-Quik (Dade Behring, Deerfield, IL, USA).
Cells were detached with a tissue culture cell scraper (Fisher, 08-100-240) and mechanicalysedysed by passing through a 27½ gauge needle.
