Microfluidic systems and methods of transport and lysis of cells and analysis of cell lysate
AssigneeUT-Battelle, LLC
InventorsChristopher T. Culbertson, Stephen C. Jacobson, Maxine A. McClain, J. Michael Ramsey
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Abstract
Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.
Claims43
A method of releasing the intracellular contents of at least one cell of a cell-containing fluid sample for analysis, said method comprising the steps of:
- a. providing a substrate having a microchannel structure which includes at least one microchannel therein;
- b. generating an electric field from a source of electric potential, said electric field being applied in a spatially defined region of said at least one microchannel, comprising a cell lysis region, and having sufficient strength to induce cell lysis;
- c. causing said cell-containing fluid sample to flow through said cell lysis region, exposing said at least one cell of said flowing fluid sample to said electric field for a time sufficient to release said intracellular contents of said at least one cell into said fluid sample, thereby providing a volume of analyte in said at least one microchannel;
- and analyzing said volume of analyte in said microchannel structure.
A microfluidic system for transport and lysis of at least one cell of a cell-containing fluid sample, said system comprising a source of electric potential, a solid substrate having at least, one microchannel with a longitudinal axis and transporting means for causing flow of a cell-containing fluid sample through said at least one microchannel, said microchannel having a first wall portion on one side of said axis, a second wall portion on another side of said axis and a cell lysis region between said first and second wall portions, a first and a second electrical contact positioned adjacent said first and second wall portions of said at least one microchannel, said first and second electrical contacts being spatially separated by said cell lysis region and being electrically isolated from one another, said first and second electrical contacts being connected to said source of electrical potential which is operative to apply an electric field to said flowing cell-containing fluid sample in said cell lysis region within the microchannel space between said first and second electrical contacts.
A microfluidic system for transport and lysis of at least one cell of a cell-containing fluid sample and separation of the intracellular content of said at least one cell, said system comprising a solid substrate having at least one microchannel disposed therein and means for causing flow of a cell-containing fluid sample through said at least one microchannel, said microchannel having a cell transport segment and a separation segment having first and second end portions, first and second electrical contacts adjacent said microchannel, intermediate said transport segment and said separation segment, and spatially separated from one another, the microchannel space between said electrical contacts defining a cell lysis region, said electrical contacts being connected to a source of electric potential operative to apply an electric field to said flowing cell-containing fluid sample in said cell lysis region;
- and means between said first and second separation segment end portions for effecting separation of said intracellular contents of said at least one cell.
A method of releasing and analyzing compounds from at least one particle of a particle-containing fluid sample, comprising the steps of:
- a. providing a substrate having a microchannel structure which includes a first microchannel intersecting with a second microchannel at an intersection;
- b. positioning a particle of a particle-containing fluid sample in said intersection, said particle containing at least one compound;
- c. introducing a chemical lysing solution into said intersection through said second microchannel, whereby said at least one compound is released into the fluid sample, thereby providing a volume of analyte in said intersection;
- d. causing said fluid sample containing said analyte to flow through and beyond said intersection;
- and then
- e. analyzing said volume of analyte in said first microchannel beyond said intersection.
Dependencies are inferred from the claim text.