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Tumor cells cultured within N3F-derived matrices were fixed in 4% paraformaldehyde, permeabilized with 100 µM digitonin, and stained for β1 integrin, E-cadherin, and F-actin (phalloidin conjugated to the tetramethyl rhodamine, Invitrogen).
After 24 h, cells and matrices were fixed and analysed by confocal microscopy.
To quantify the degree of capillary formation, 3D matrices were fixed at 24, 36, 48, 60, and 72 h by aspirating the medium, washing with PBS, and then fixing with 4% glutaraldehyde in PBS, pH 7.4, overnight at 4°C.
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The covariance matrices are fixed, since in this case the NN weights are frozen.
Following the terminology of control engineering, linear parameter-varying (LPV) systems are time-varying plants whose state space matrices are fixed functions of some vector of varying parameters.
In fact, when the modes in the connection weight matrices are fixed, two additive time-varying delay components may also has finite modes, and the switching between different modes can also be governed by a Markov chain.
In fact, when the modes in the connection weight matrices are fixed, two additive time-varying delay components may also have finite modes due to the dynamic systems subject to abrupt variation frequently in their structures, and the switching between different modes can also be governed by a Markov chain.
At the end of each culture period, the medium was removed and the cells and matrix were fixed with HistoChoice MB (Amresco, Solon, OH, USA).
Invaded cells on the lower surface of the Matrigel matrix were fixed with 4% PFA and stained with 0.2% crystal violet.
It is assumed that the demand matrix is fixed and given a priori.
In the simulations, the unknown combining matrix was fixed and was taken as H = 1 0.3 0.3 1.
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