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n Selecting a background color: To select a new background color, Alt-click (Option-click on the Mac) the desired color with the Eyedropper tool. (Again, this assumes that the foreground icon is selected in the Color palette. If the background icon is selected, click with the Eyedropper to lift the background color.) n Skating over the color spectrum: You can animate the foreground color icon by clicking and dragging with the Eyedropper tool. As soon as you achieve the desired color, release the mouse button. To animate the background color icon, Alt-drag (Option-drag on the Mac) with the Eyedropper tool. The icon color changes as you move the Eyedropper tool. Again, swap these procedures if the background color icon is selected in the Color palette.



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The above process is adaptable for use with many different cell types However, the procedure will vary with the characteristics of the desired protein In our case, collagen type I retained its bioactivity for hepatocyte attachment and spreading despite treatment with acetone, ethanol, and dehydration Other proteins may require modi cations of this protocol to retain their bioactivity Additionally, in our experience, covalent binding of collagen is not required for hepatocyte patterning and simple adsorption to the glass substrate is adequate Yet it should be noted that reliance on adsorption alone is likely to result in different immobilized protein conformations than covalently bound proteins, and these potential differences should be kept in mind if modifying this procedure for other adhesive molecules Furthermore, selection of medium for two cell types must be considered Medium must be selected that provides adequate nutrients and buffering for each population.





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(b) Use Remark 6.1 to conclude that the desired optimal solution, wo, the constrained optiof mization problem is given by

Also, because of the rapid mitotic rate of broblasts, our medium contains hydrocortisone as a growth inhibitor Alternatively, as suggested above, under some circumstances broblasts can be chemically growth arrested with mitomycin C Potential problems include minimal cell adhesion to the substrate and, conversely, uniform cell adhesion to the substrate Lack of cell adhesion to the substrate can indicate many problems The most common, however, is underdevelopment of the exposed photoresist (See Protocol 158, Step (e)), resulting in a lack of exposed borosilicate for protein immobilization This can be alleviated by increasing development time, or increasing exposure to oxygen plasma before surface modi cation This effect is often exacerbated with small (<5 m) pattern dimensions Alternatively, contaminants that coat the glass and prevent protein immobilization, or commonly utilized undercoatings for promoting photoresist adhesion, could produce this effect.

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There are many variables in the Figure 7.14 Lines, Symbols and Arrows menu. Pick the style you want, but don't change any of the default sizes because that will affect your dimensions unless you really know what you are doing. Change the sizes in the Fit menu. See Tutorial 7a and 7b for how to do this.

On the other hand, a lack of discernible pattern because of exuberant cell adhesion is frequently caused by a defect in the original photoresist coating Photoresist can degrade and crack over time, allowing exposure of all areas of the substrate to surface modi cation Also, excessive exposure to oxygen plasma will strip the photoresist from the surface of the wafer completely, allowing for homogeneous protein adsorption In summary, the above protocols describe a method for micropatterning hepatocyte- broblast cocultures based on standard microfabrication techniques Utilization of this system enables the controlled investigation of the complex cellcell interactions important in the stabilization of the hepatocyte phenotype..

Figure 9-22

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