Cell analysis methods
First Claim
1. A method of analyzing cells in a carrier solution, comprising:
- (a) introducing said carrier solution into a conduit having a surface portion wherein the surface portion has a discreet capture zone formed therein, said carrier solution having a plurality of cells suspended therein;
(b) contacting said plurality of said cells to said surface portion, said surface portion containing at least one imaging field;
(c) sequentially interrogating said plurality of cells in said imaging field with at least two different types of emitted light;
(d) processing resultant light from said imaging field for each of said at least two different types of emitted light;
(e) generating digital information for each of said plurality of cells from said resultant light for each of said at least two different types of emitted light; and
then (f) generating a response file for each of said plurality of cells from said digital information for each of said at least two different types of emitted light;
and wherein said sequentially interrogating step is followed by the steps of;
altering the temperature of said cells in said capture zone; and
then repeating said sequentially interrogating step.
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Accused Products
Abstract
A method of analyzing cells in a carrier solution comprises the following steps: (a) Introducing the carrier solution into a conduit having a surface portion (preferably a substantially flat surface portion). The carrier solution has the cells suspended therein. (b) Allowing the cells to settle on the surface portion, the surface portion including at least one imaging field. In an alternate embodiment, one or more discreet capture zones (e.g., formed from an affinity species immobilized on the substrate or a textured region on the substrate) are formed on the surface portion, and this step (b) comprises capturing the cells in the capture zone. (c) Sequentially interrogating a plurality of the cells in the imaging field with emitted light. (d) Processing resultant light from the imaging field. (e) Generating digital information for each of the plurality of cells from the resultant light. (f) Generating a response file for each of the plurality of cells from the digital information. The response file generated in the method can be used for the sizing, enumeration, characterization, and/or classification of cells in the sample. Apparatus for carrying out the foregoing method is also disclosed, along with methods and apparatus employing reciprocal flow techniques.
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Citations
31 Claims
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1. A method of analyzing cells in a carrier solution, comprising:
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(a) introducing said carrier solution into a conduit having a surface portion wherein the surface portion has a discreet capture zone formed therein, said carrier solution having a plurality of cells suspended therein;
(b) contacting said plurality of said cells to said surface portion, said surface portion containing at least one imaging field;
(c) sequentially interrogating said plurality of cells in said imaging field with at least two different types of emitted light;
(d) processing resultant light from said imaging field for each of said at least two different types of emitted light;
(e) generating digital information for each of said plurality of cells from said resultant light for each of said at least two different types of emitted light; and
then(f) generating a response file for each of said plurality of cells from said digital information for each of said at least two different types of emitted light;
and wherein said sequentially interrogating step is followed by the steps of;
altering the temperature of said cells in said capture zone; and
then repeating said sequentially interrogating step.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
and wherein said sequentially interrogating step is repeated for each of said discrete capture zones.
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24. The method according to claim 4, wherein said cells are blood cells, and wherein said blood cells bind to said capture zone.
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25. The method according to claim 1, wherein the flow of said cells in said solution during said contacting step is modified by feedback from said resultant light or said response files.
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26. The method according to claim 1, wherein said sequentially interrogating step is followed by altering the rate of flow of said solution through said conduit;
- and then repeating said sequentially interrogating step.
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27. The method according to claim 1, further comprising lysing cells bound to said capture zone, and then analyzing nucleic acid released from said lysed cells;
and wherein said lysing follows said contacting step.
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28. The method according to claim 1, further comprising transiently permeabilizing said cells to release a portion of the contents thereof, while retaining nucleic acid for subsequent analysis therein;
and wherein said cells are transiently permeabilized after said contacting step.
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29. The method according to claim 4, wherein said capture zone comprises an affinity species immobilized on said surface portion.
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30. The method according to claim 4, wherein said capture zone comprises a textured segment of said surface portion.
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31. The method according to claim 1, further comprising permeabilizing cells bound to said first capture zone;
and wherein said permeabilizing step follows said contacting step.
Specification