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Using both models, time-varying heat transfer within the ice medium as well as the ground is investigated.
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Sure enough, the photos showed sharp delineations between bright dry ice, medium-bright water ice, and dark rock.
Immediately after irradiation (20 Gy) on ice, the medium was replaced with warm medium and cells were incubated (37°C) for repair.
Afterwards, cells were washed twice with ice-cold medium, detached with ice-cold PBS and washed twice by centrifugation.
Cells cultured for 9 days were washed twice with ice-cold medium, fixed by ice-cold methanol, and stained with 0.2% crystal violet.
Incorporation of CFSE dye was stopped by adding 5 volumes of ice-cold medium and incubation on ice for 5 minutes.
The tissues were stored in ice cold medium DMEM/F12 supplemented with 10% fetal bovine serum (FBS), penicillin and streptomycin.
Cells were centrifuged for 5 minutes, washed in ice cold medium, and resuspended in lysis buffer as described in [ 24].
Immediately after the end of the treatment, the plates were put on ice, the medium was taken off and 1 ml of 4% paraformaldehyde was added per well.
Briefly, the treated cells were placed on ice, the medium was removed, and the cells were washed once with cold PBS.
Monolayers were rapidly cooled by placing on ice, changing medium to phosphate buffered saline with 0.5 mg/ml sulfo-NHS biotin only on the apical side.
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