Methods and systems for releasing intracellular material from cells within microfluidic samples of fluids
First Claim
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1. A microfluidic device comprising:
- a lysing module configured to receive a cell-containing microdroplet;
an actuator located upstream of the lysing module and configured to create a difference between an upstream pressure and a downstream pressure acting on the cell-containing microdroplet to move the microdroplet towards the lysing module;
a vented positioning element located upstream of the lysing module and downstream of the actuator, wherein the vented positioning element is configured to stop the cell-containing microdroplet in a lysing position with respect to the lysing module, and wherein the vented positioning element is configured to position a portion of the cell-containing microdroplet downstream of the vented positioning element in the lysing position; and
a lysing mechanism within the lysing module, configured to release intracellular material from cells within the cell-containing microdroplet in the lysing position with respect to the lysing module.
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Abstract
The present invention relates to a microfluidic system for processing a cell-containing liquid. The system includes a lysing zone to receive the cell-containing sample and a positioning element to position the cell-containing sample in a lysing position in the vicinity of a lysing mechanism. The lysing mechanism releases intracellular material, such as DNA or RNA, from the cells. In one embodiment, the lysing mechanism includes electrodes for generating an electric field sufficient to release intracellular contents from the cells. Alternatively, the lysing mechanism may lyse the cells using chemical, heat and/or ultrasonic techniques or any combination of these techniques.
379 Citations
31 Claims
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1. A microfluidic device comprising:
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a lysing module configured to receive a cell-containing microdroplet; an actuator located upstream of the lysing module and configured to create a difference between an upstream pressure and a downstream pressure acting on the cell-containing microdroplet to move the microdroplet towards the lysing module; a vented positioning element located upstream of the lysing module and downstream of the actuator, wherein the vented positioning element is configured to stop the cell-containing microdroplet in a lysing position with respect to the lysing module, and wherein the vented positioning element is configured to position a portion of the cell-containing microdroplet downstream of the vented positioning element in the lysing position; and a lysing mechanism within the lysing module, configured to release intracellular material from cells within the cell-containing microdroplet in the lysing position with respect to the lysing module. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A microfluidic device, comprising:
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a lysing module configured to receive a microdroplet of cell-containing sample; a lysing mechanism within the lysing module, configured to release intracellular contents from cells in the microdroplet of cell-containing sample within the lysing module; a first gas actuator situated upstream of the lysing module and configured to move the microdroplet of cell-containing sample downstream to overlap the lysing module; a positioning element located downstream of the lysing module and configured to inhibit downstream movement of the cell containing sample, thereby positioning at least some of the cell containing sample in a lysing position with respect to the lysing module; and a second gas actuator disposed upstream from the lysing module but downstream from the first actuator, to provide a gas pressure sufficient to;
(a) prepare a lysed microdroplet comprising intracellular contents released from cells of the cell-containing sample within the lysing module, the microdroplet having a length equal to a distance between the second gas actuator and the positioning element and (b) move the lysed microdroplet downstream of the lysing module and past the positioning element. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23)
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24. A method for lysing a microdroplet of cell-containing liquid, comprising:
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introducing the microdroplet of cell-containing liquid to a lysing module of a microfluidic device; inhibiting the microdroplet of cell-containing liquid from moving downstream from the lysing module; actuating a lysing mechanism to release intracellular contents from cells of the cell-containing liquid within the lysing module; and providing a gas pressure sufficient to separate a first portion of the microdroplet of cell-containing liquid located within the lysing module from a second portion of the microdroplet of cell-containing liquid located upstream of the lysing module thereby preparing a lysed microdroplet comprising intracellular contents released from cells of the cell-containing liquid within the lysing module. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31)
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Specification