Microfludic protein crystallography techniques
First Claim
1. A method for mixing a first fluid and a second fluid in different portions in a microfluidic device, the method comprising:
- providing a microfluidic device comprising a plurality of compound wells, wherein each compound well comprises a first well having a first volume and a second well having a second volume, wherein a ratio of the first volume to the second volume of at least one compound well differs from that of the remaining compound wells;
filling the first wells with said first fluid;
filling the second wells with said second fluid; and
placing the first well of each compound well in fluid communication with the second well, thereby allowing the first fluid and second fluid to mix by free-interface diffusion in portions that differ between compound wells.
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Abstract
The present invention relates to microfluidic devices and methods facilitating the growth and analysis of crystallized materials such as proteins. In accordance with one embodiment, a crystal growth architecture is separated by a permeable membrane from an adjacent well having a much larger volume. The well may be configured to contain a fluid having an identity and concentration similar to the solvent and crystallizing agent employed in crystal growth, with diffusion across the membrane stabilizing that process. Alternatively, the well may be configured to contain a fluid having an identity calculated to affect the crystallization process. In accordance with the still other embodiment, the well may be configured to contain a material such as a cryo-protectant, which is useful in protecting the crystalline material once formed.
212 Citations
22 Claims
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1. A method for mixing a first fluid and a second fluid in different portions in a microfluidic device, the method comprising:
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providing a microfluidic device comprising a plurality of compound wells, wherein each compound well comprises a first well having a first volume and a second well having a second volume, wherein a ratio of the first volume to the second volume of at least one compound well differs from that of the remaining compound wells; filling the first wells with said first fluid; filling the second wells with said second fluid; and placing the first well of each compound well in fluid communication with the second well, thereby allowing the first fluid and second fluid to mix by free-interface diffusion in portions that differ between compound wells. - View Dependent Claims (2, 3, 4, 5)
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6. A method of mixing a first fluid and a second fluid in adjustable portions in a microfluidic device, the method comprising:
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providing a microfluidic device comprising a first well, a plurality of first control channels disposed across the first well, and a second well; filling the first well with said first fluid; filling the second well with said second fluid; pressurizing at least one first control channel to divide the first well into a plurality of first chambers; and placing the first well in fluid communication with the second well, such that at least one of said first chambers is in fluid communication with the second well, thereby allowing the first fluid and second fluid to mix by free-interface diffusion in portions determined by the pressurized first control channels. - View Dependent Claims (7, 8, 9, 10, 11)
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12. A method of mixing a first fluid and a second fluid at varying rates in a microfluidic device, the method comprising:
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providing a microfluidic device comprising a plurality of compound wells, wherein each compound well comprises a first well, a second well, and one or more microchannels connecting the first and second wells, wherein the amount, average length, and/or average cross sectional area of the microchannels vary between compound wells; filling the first wells with said first fluid; filling the second wells with said second fluid; and placing the first wells in fluid communication with the second wells via the microchannels, thereby allowing the first fluid and second fluid to mix by free-interface diffusion at rates that vary between compound wells. - View Dependent Claims (13, 14, 15, 16)
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17. A method of capturing, in a microfluidic device, a chemical gradient resulting from the diffusive mixing of a first fluid and a second fluid, the method comprising:
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providing a microfluidic device comprising a first well, at least one first control channel disposed across the first well, and a second well; filling the first well with said first fluid; filling the second well with said second fluid; placing the first well in fluid communication with the second well, thereby allowing the first fluid and second fluid to begin mixing by free-interface diffusion; and pressurizing at least one first control channel before mixing has completed, thereby dividing the first well into a plurality of first chambers, such that concentrations of the first fluid and second fluid differ between first chambers. - View Dependent Claims (18, 19, 20, 21, 22)
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Specification