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All tissues were placed in Tissue Tek OCT (Sakura Finetek, Torrance, CA) and frozen over liquid nitrogen.
After fixation, lung tissues were placed in 10% sucrose until tissues sink, then 20% overnight.
The tissues were placed in clean watch glasses and were oven dried at 105 °C for 1 h and later cooled in the desiccators.
Tissues were placed in the EPON for 2 h and then incubated with pure resin at 68°C for 48 h to complete the polymerization.
Tissues were placed in a 10% solution of buffered formalin.
The tissues were placed in 10% neutral buffered formalin for three days to re-hydrate and fix the tissue.
Immediately after dissection tissues were placed in RNAse free 1.5ml tubes (Biopur, Eppendorf) and immersed in RNAlater reagent (Qiagen) to minimize RNA degradation.
Tissues were placed in separated sterile 50-mL tubes with ice-cold culture medium and then transferred to a cell culture laboratory on ice.
In the study of CO histochemistry plus Nissl's counterstaining, the brain tissues were placed in sucrose/PBS with concentrations of 10 %, 20 and 30% and 30%ially until they sequentially
In contrast, when tissues were placed in culture for the continuous assay of bioluminescence, rhythms were observed in embryonic (E18) tissues.
Then the tissues were placed in 3% hydrogen peroxide and absolute methanol for 5 minutes to reduce endogenous peroxidase activity, followed by washing in PBS.
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