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Each cover slip of cells was assembled into a purpose-built chamber, a fresh aliquot of modified HBSS with or without 1 μM STS was added to the cells and placed on the stage of a Leica TCS SP5 II confocal laser scanning microscope (Leica Microsystems GmbH, Wetzlar, Germany).
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At the end of the respective incubation periods, the cover slips of cells were fixed with 4% formaldehyde (Formaldehyde solution min. 37% free from acid, cat#103999, 64293 Darmstadt, Germany) in PBS.
For Wnt pathway investigations, the respective cover slips of cells were incubated with either Wnt3A conditioned media (40% in DMEM + 10% FCS) or cycloheximide (CHX, 100 μg/ml in DMEM + 10% FCS) for 0, 1, 2 or 4 hours selectively at 37˚C.
And some said that thalidomide slipped into the DNA of cells in a developing limb, preventing the cells from making the proteins they needed.
Gascoigne and Taylor proposed the independent pathway model by showing that the fraction of cells that slip or die can be altered by perturbing the kinetics of either pathway [7].
The slip of both the neighboring cells must be non-zero for the smoothness constraint to be effective.
This morphology is in agreement with previous description of cells that slipped through mitosis, namely after a prolonged mitotic delay in the presence of spindle poisons [11], [12], [20].
The control cells were normal and spread on the cover slip of the well plate whereas cells shrank in the formulations-treated group.
In all cases, 100% of cells scored eventually slipped out of mitosis.
Alternatively, Perspex cover slips can be used for the collection of cells [ 12].
Each cover slip was divided into quadrants and the number of cells (total and positive for staining) were counted manually.
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CEO of Professional Science Editing for Scientists @ prosciediting.com