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This was followed by overnight hybridisation at 37°C in a humid chamber and washing of tissue sections for 3 × 5 min in 0.1X SSC (Saline-Sodium-Citrate) at 60°C and mounting with 4'-6-Diamidino-2-phenylindole (DAPI) counterstaining.
The perforated tubes with the centrifuged leaf strips inside were brought back to the lab in a humid box and the new tissue weight was measured prior to the next round of vacuum infiltration.
Primary antibodies were reacted with the tissue overnight in a humid chamber at 4 °C and washed with PBS.
The tissues were incubated in a humid 5% CO2 incubator at 37°C to allow the attachment of the tissue to the surface of the flask.
The slides with released pollen grains were incubated upside down for 6 hours to overnight in a humid chamber made from tip boxes, wet tissue paper, and two pasteur pipettes to hold the slides.
Then, the deposit was resuspended in culture medium and plated in 10 cm tissue culture flasks and maintained in humid incubator at 37 °C and 5% CO2.
Cells were cultured at 37°C with 5% CO2 in a humid atmosphere in T75 flasks or 24-well tissue culture plates (Falcon; Becton Dickinson).
Needs high humidity ( 60 - 80'F humid ).
The tissue pieces were subsequently incubated at 37 °C in humid air with 5% CO2, and the medium was replaced every 3 days after the initial plating.
Fifty microliters of synchronized iRBC suspension, at the concentration of 108/ml, were overlaid onto each tissue section for 60 minutes at 37°C in a humid chamber.
Muscle tissue was kept at −16 °C and in a 100%% humid medium.
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