Method of making device for arraying biomolecules and for monitoring cell motility in real-time
View Patent ↗The invention relates to devices, devices for arraying biomolecules, including cells, methods for arraying biomolecules, assays for monitoring cellular movement, and systems for monitoring cellular movement. The devices include a support; a first layer configured to be placed in fluid-tight contact with the support, the first layer having an upper surface and defining a pattern of micro-orifices, each micro-orifice of the pattern of micro-orifices having walls and defining a micro-region on the support when the first layer is placed in fluid-tight contact with the support such that the walls of said each micro-orifice and the micro-region on the support together define a micro-well; and a second layer configured to be placed in fluid-tight contact with the upper surface of the first layer, the second layer defining a pattern of macro-orifices, each macro-orifice of the pattern of macro-orifices having walls and defining a macro-region when the first layer is placed in fluid-tight contact with the support and the second layer is placed in fluid-tight contact with the first layer such that the walls of the macro-orifice and the macro-region together define a macro-well.
1. A method of fabricating a device comprising:
a. providing a support;
b. forming a first layer configured to be placed in fluid-tight contact with the support, the first layer having an upper surface and defining a pattern of micro-orifices, each micro-orifice of the pattern of micro-orifices having walls and defining a micro-region on the support when the first layer is placed in fluid-tight contact with the support such that the walls of said each micro-orifice and the micro-region on the support together define a micro-well, wherein said first layer is formed by:
providing a first mold having a pattern of micro-posts corresponding to the pattern of micro-orifices;
applying a first elastomeric material in liquid form to the first mold;
curing the first elastomeric material; and
removing the cured first elastomeric material from the first mold;
c. forming a second layer configured to be placed in fluid-tight contact with the upper surface of the first layer, the second layer defining a pattern of macro-orifices, each macro-orifice of the pattern of macro-orifices having walls and defining a macro-region when the first layer is placed in fluid-tight contact with the support and the second layer is placed in fluid-tight contact with the first layer such that the walls of the macro-orifice and the macro-region together define a macro-well, wherein said second layer is formed by:
providing a second mold having a pattern of macro-posts corresponding to the pattern of macro-orifices;
applying a second elastomeric material in liquid form to the second mold;
curing the second elastomeric material; and
removing the cured second elastomeric material from the second mold; and
d. placing the first layer in fluid-tight contact with the support.
2. The method of claim 1 , wherein forming a first layer comprises providing a first layer configured to be placed in conformal contact with the support when the first layer is placed against the support.
3. The method of claim 1 , wherein forming a second layer comprises providing a second layer configured to be placed in conformal contact with the first layer when the second layer is placed against the first layer.
4. The method of claim 1 , wherein forming a first layer comprises forming a first layer made of polydimethylsiloxane.
5. The method of claim 1 , wherein forming a second layer comprises forming a second layer made of polydimethylsiloxane.
6. The method of claim 1 , further comprising placing the second layer against the first layer such that each macro-region encompasses at least one micro-region.
7. The method of claim 6 , wherein placing comprises placing the second layer against the first layer such that each macro-region encompasses a plurality of micro-regions.
8. The method of claim 1 , wherein applying an elastomeric material includes spin-coating the elastomeric material.
9. The method of claim 1 , further comprising providing an adhesive adapted to be applied between the first layer and the second layer when the second layer is placed against the first layer.