U.S. patent application number 14/806124 was filed with the patent office on 2015-11-26 for method and apparatus for programmable fluidic processing.
The applicant listed for this patent is Board of Regents, The University of Texas System. Invention is credited to Frederick F. BECKER, Giovanni De GASPERIS, Peter GASCOYNE, Jody Valentine VYKOUKAL, Xiaobo WANG.
Application Number | 20150336111 14/806124 |
Document ID | / |
Family ID | 22945720 |
Filed Date | 2015-11-26 |
United States Patent
Application |
20150336111 |
Kind Code |
A1 |
BECKER; Frederick F. ; et
al. |
November 26, 2015 |
Method and Apparatus for Programmable Fluidic Processing
Abstract
A method and apparatus for microfluidic processing by
programmably manipulating a packet. A material is introduced onto a
reaction surface and compartmentalized to form a packet. A position
of the packet is sensed with a position sensor. A programmable
manipulation force is applied to the packet at the position. The
programmable manipulation force is adjustable according to packet
position by a controller. The packet is programmably moved
according to the programmable manipulation force along arbitrarily
chosen paths.
Inventors: |
BECKER; Frederick F.;
(Houston, TX) ; GASCOYNE; Peter; (Bellaire,
TX) ; WANG; Xiaobo; (Sugarland, TX) ;
VYKOUKAL; Jody Valentine; (Houston, TX) ; De
GASPERIS; Giovanni; (L'Aquila, IT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Board of Regents, The University of Texas System |
Austin |
TX |
US |
|
|
Family ID: |
22945720 |
Appl. No.: |
14/806124 |
Filed: |
July 22, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14452047 |
Aug 5, 2014 |
|
|
|
14806124 |
|
|
|
|
13545775 |
Jul 10, 2012 |
8834810 |
|
|
14452047 |
|
|
|
|
12622775 |
Nov 20, 2009 |
8216513 |
|
|
13545775 |
|
|
|
|
11135615 |
May 23, 2005 |
7641779 |
|
|
12622775 |
|
|
|
|
09902933 |
Jul 10, 2001 |
6977033 |
|
|
11135615 |
|
|
|
|
09249955 |
Feb 12, 1999 |
6294063 |
|
|
09902933 |
|
|
|
|
Current U.S.
Class: |
204/547 ;
204/643 |
Current CPC
Class: |
B01L 2200/143 20130101;
G01N 27/44773 20130101; B01L 2400/0454 20130101; B01F 13/0076
20130101; B01L 3/502715 20130101; B01L 3/502761 20130101; B01L
2400/0421 20130101; B03C 5/005 20130101; B01L 2300/089 20130101;
B03C 5/026 20130101; G01N 27/447 20130101; B03C 5/024 20130101;
B01L 3/502792 20130101; B01L 2400/0424 20130101; B01F 2215/0404
20130101; B03C 5/028 20130101 |
International
Class: |
B03C 5/00 20060101
B03C005/00; B03C 5/02 20060101 B03C005/02; B01L 3/00 20060101
B01L003/00; G01N 27/447 20060101 G01N027/447 |
Claims
1-41. (canceled)
42. A device to manipulate at least one packet, comprising: a
reaction surface; an array of electrodes operably coupled to the
reaction surface and adapted to generate electrical field
distributions to impart manipulation forces to move at least one
packet disposed proximate the reaction surface to different
positions about the reaction surface; and at least one position
sensor configured to monitor a position of the at least one
packet.
43. The device of claim 42, wherein the at least one position
sensor comprises a plurality of position sensors.
44. The device of claim 43, wherein the plurality of position
sensors are adapted to track the position of the at least one
packet.
45. The device of claim 42, wherein the at least one position
sensor is separate from the array of electrodes.
46. The device of claim 42, wherein the reaction surface is an
integrated circuit.
47. The device of claim 42, wherein the array of electrodes is a
two dimensional array of electrodes.
48. The device of claim 42, further comprising at least one of a
dielectric coating or a hydrophobic coating covering the array of
electrodes.
49. The device of claim 42, further comprising a partitioning
medium in which the at least one packet can be suspended.
50. The device of claim 42, further comprising a fluidic sensor to
measure an additional property of the at least one packet.
51. The device of claim 50, wherein the fluidic sensor comprises at
least one of an optical microscope, a chemical sensor, an
electrochemical sensor, an electrical sensor, or an optical
sensor.
52. The device of claim 42, further comprising a capillary in
communication with the reaction surface.
53. The device of claim 42, wherein the array of electrodes is
adapted to adjust the manipulation forces based on the position of
the at least one packet as monitored by the at least one position
sensor.
54. The device of claim 42, wherein the at least one packet is a
fluid packet.
55. A system to manipulate at least one packet, comprising: a
device comprising a reaction surface, an array of electrodes
operably coupled to the reaction surface and adapted to generate
electrical field distributions to impart manipulation forces to
move at least one packet disposed proximate the reaction surface to
different positions about the reaction surface, and at least one
position sensor configured to monitor a position of the at least
one packet; and a controller coupled to the array of electrodes and
the at least one position sensor, the controller adapted to address
the array of electrodes to generate the electrical field
distributions and thereby impart the manipulation forces on the at
least one packet to move the at least one packet to different
positions about the reaction surface.
56. The system of claim 55, wherein the at least one position
sensor comprises a plurality of position sensors.
57. The system of claim 56, wherein the plurality of position
sensors are adapted to track the position of the at least one
packet.
58. The system of claim 55, wherein the at least one position
sensor is separate from the array of electrodes.
59. The system of claim 55, wherein the at least one packet is a
fluid packet.
60. The system of claim 55, wherein the reaction surface is an
integrated circuit
61. The system of claim 55, wherein the array of electrodes is a
two dimensional array of electrodes.
62. The system of claim 55, wherein the device further comprises at
least one of a dielectric coating or a hydrophobic coating covering
the array of electrodes.
63. The system of claim 55, wherein the device further comprises a
partitioning medium in which the at least one packet can be
suspended.
64. The system of claim 55, wherein the device further comprises a
fluidic sensor to measure an additional property of the at least
one packet.
65. The system of claim 64, wherein the fluidic sensor comprises at
least one of an optical microscope, a chemical sensor, an
electrochemical sensor, an electrical sensor, or an optical
sensor.
66. The system of claim 55, wherein the device further comprises a
capillary in communication with the reaction surface.
67. The system of claim 55, wherein the controller is adapted to
adjust the manipulation forces based on the position of the at
least one packet as monitored by the at least one position
sensor.
68. The system of claim 67, wherein the controller is adapted to
adjust manipulation forces based on the position of the at least
one packet to achieve movement of the at least one packet along a
desired path.
69. The system of claim 67, wherein the controller is adapted to
adjust manipulation forces based on the position of the at least
one packet to merge the at least one packet with at least one
additional packet.
70. The system of claim 67, wherein the controller is adapted to
adjust manipulation forces based on the position of the at least
one packet to perform at least one function selected from mixing or
splitting the at least one packet.
71. A method to manipulate at least one packet, comprising:
providing a device comprising a reaction surface, and an array of
electrodes operably coupled to the reaction surface and adapted to
generate electrical field distributions to impart manipulation
forces to move at least one packet disposed proximate the reaction
surface to different positions about the reaction surface;
disposing at least one packet proximate the reaction surface;
generating electrical field distributions by addressing the array
of electrodes to impart manipulation forces on the at least one
packet to move the at least one packet to different positions about
the reaction surface; and monitoring a position of the at least one
packet about the reaction surface.
72. The method of claim 71, wherein monitoring the position
includes providing at least one position sensor and monitoring the
position of the at least one packet by the at least one position
sensor.
73. The method of claim 72, wherein the at least one position
sensor comprises a plurality of position sensors.
74. The method of claim 73, wherein monitoring the position
comprises tracking the position of the at least one packet by the
plurality of position sensors.
75. The method of claim 72, wherein the at least one position
sensor is separate from the array of electrodes.
76. The method of claim 71, wherein the reaction surface is an
integrated circuit
77. The method of claim 71, wherein the array of electrodes is a
two dimensional array of electrodes.
78. The method of claim 71, wherein the device further comprises a
partitioning medium in which the at least one packet can be
suspended.
79. The method of claim 71, wherein the device further comprises a
fluidic sensor to measure an additional property of the at least
one packet.
80. The method of claim 79, wherein the fluidic sensor comprises at
least one of an optical microscope, a chemical sensor, an
electrochemical sensor, an electrical sensor, or an optical
sensor.
81. The method of claim 71, wherein the device further comprises a
capillary in communication with the reaction surface.
82. The method of claim 71, further comprising adjusting the
manipulation forces based on the position of the at least one
packet as monitored by the at least one position sensor.
83. The method of claim 71, wherein the at least one packet is a
fluid packet.
Description
[0001] This application is a continuation of co-pending U.S.
application Ser. No. 14/452,047, filed Aug. 5, 2014, which is a
continuation of U.S. application Ser. No. 13/545,775 filed Jul. 10,
2012 (and now issued as U.S. Pat. No. 8,834,810), which is a
continuation of co-pending U.S. application Ser. No. 12/622,775
filed Nov. 20, 2009 (and now issued as U.S. Pat. No. 8,216,513),
which is a continuation of U.S. application Ser. No. 11/135,615
filed May 23, 2005 (and now issued as U.S. Pat. No. 7,641,779),
which is a continuation of U.S. application Ser. No. 09/902,933
filed Jul. 10, 2001 (and now issued as U.S. Pat. No. 6,977,033),
which is a continuation of U.S. application Ser. No. 09/249,955,
filed Feb. 12, 1999 (and now issued as U.S. Pat. No. 6,294,063).
The entire text of each of the above-referenced disclosures is
specifically incorporated by reference herein without
disclaimer.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates generally to fluidic
processing and, more particularly, to a method and apparatus for
programmably manipulating and interacting one or more
compartmentalized packets of material on a reaction surface.
[0004] 2. Description of Related Art
[0005] Chemical protocols often involve a number of processing
steps including metering, mixing, transporting, division, and other
manipulation of fluids. For example, fluids are often prepared in
test tubes, metered out using pipettes, transported into different
test tubes, and mixed with other fluids to promote one or more
reactions. During such procedures, reagents, intermediates, and/or
final reaction products may be monitored, measured, or sensed in
analytical apparatus. Microfluidic processing generally involves
such processing and monitoring using minute quantities of fluid.
Microfluidic processing finds applications in vast fields of study
and industry including, for instance, diagnostic medicine,
environmental testing, agriculture, chemical and biological warfare
detection, space medicine, molecular biology, chemistry,
biochemistry, food science, clinical studies, and pharmaceutical
pursuits.
[0006] A current approach to fluidic and microfluidic processing
utilizes a number of microfluidic channels that are configured with
microvalves, pumps, connectors, mixers, and detectors. While
devices using micro-scale implementations of these traditional
approaches may exhibit at least a degree of utility, vast room for
improvement remains. For instance, pumps and valves used in
traditional fluidic transportation are mechanical. Mechanical
devices, particularly when coupled to thin microchannels, may be
prone to failure or blockage. In particular, thin channels may
become narrowed or partially-blocked due to buildup of channel
contamination, which, in turn, may lead to mechanical failure of
associated devices. Current microfluidic devices also lack
flexibility, for they rely upon a fixed pathway of microchannels.
With fixed pathways, devices are limited in the number and type of
tasks they may perform. Also, using fixed pathways makes many types
of metering, transport, and manipulation difficult. With
traditional devices, it is difficult to partition one type of
sample from another within a channel.
[0007] Electrical properties of materials have been employed to
perform a limited number of fluidic processing tasks. For example,
dielectrophoresis has been utilized to aid in the characterization
and separation of particles, including biological cells. An example
of such a device is described in U.S. Pat. No. 5,344,535 to Betts,
incorporated herein by reference. Betts establishes
dielectrophoretic collection rates and collection rate spectra for
dielectrically polarizable particles in a suspension. Particle
concentrations at a certain location downstream of an electrode
structure are measured using a light source and a light detector,
which measures the increased or decreased absorption or scattering
of the light which, in turn, indicates an increase or decrease in
the concentration of particles suspended in the fluid. Although
useful for determining particle dielectrophoretic properties, such
a system is limited in application. In particular, such a system
does not allow for general fluidic processing involving various
interactions, sometimes performed simultaneously, such as metering,
mixing, fusing, transporting, division, and general manipulation of
multiple reagents and reaction products.
[0008] Another example of using certain electrical properties for
specific types of processing is disclosed in U.S. Pat. No.
5,632,957 to Heller et al., incorporated herein by reference.
There, controlled hybridization may be achieved using a matrix or
array of electronically addressable microlocations in conjunction
with a permeation layer, an attachment region and a reservoir. An
activated microlocation attracts charged binding entities towards
an electrode. When the binding entity contacts the attachment
layer, which is situated upon the permeation layer, the
functionalized specific binding entity becomes covalently attached
to the attachment layer. Although useful for specific tasks such as
DNA hybridization, room for improvement remains. In particular,
such a system, utilizing attachment sites for certain binding
entities is designed for particular applications and not for
general fluidic processing of a variety of fluids. More
specifically, such a system is designed for use with charged
binding entities that interact with attachment sites.
[0009] Another example of processing is disclosed in U.S. Pat. No.
5,126,022 to Soane et al., incorporated herein by reference. There,
charged molecules may be moved through a medium that fills a trench
in response to electric fields generated by electrodes. Although
useful for tasks such as separation, room for improvement remains
in that such devices are not well suited for performing a wide
variety of fluidic processing interactions on a wide variety of
different materials.
[0010] There are other examples of using dielectrophoresis for
performing specific, limited fluidic processing tasks. U.S. Pat.
No. 5,795,457 to Pethig and Burt, incorporated herein by reference,
disclose a method for promoting reactions between particles
suspended in liquid by applying two or more electrical fields of
different frequencies to electrode arrays. While perhaps useful for
facilitating certain interactions between many particles of
different types, the method is not well suited for general fluidic
processing. U.S. Pat. No. 4,390,403 to Batchelder, incorporated
herein by reference, discloses a method and apparatus for
manipulation of chemical species by dielectrophoretic forces.
Although useful for inducing certain chemical reactions, its
flexibility is limited, and it does not allow for general,
programmable fluidic processing.
[0011] Any problems or shortcomings enumerated in the foregoing are
not intended to be exhaustive but rather are among many that tend
to impair the effectiveness of previously known processing
techniques. Other noteworthy problems may also exist; however,
those presented above should be sufficient to demonstrated that
apparatus and methods appearing in the art have not been altogether
satisfactory.
SUMMARY OF THE INVENTION
[0012] In one respect, the invention is an apparatus for
programmably manipulating a packet. As used herein, "packet" refers
to compartmentalized matter and may refer to a fluid packet, an
encapsulated packet, and/or a solid packet. A fluid packet refers
to one or more packets of liquids or gases. A fluid packet may
refer to a droplet or bubble of a liquid or gas. A fluid packet may
refer to a droplet of water, a droplet of reagent, a droplet of
solvent, a droplet of solution, a droplet of sample, a particle or
cell suspension, a droplet of an intermediate product, a droplet of
a final reaction product, or a droplet of any material. An example
of a fluid packet is a droplet of aqueous solution suspended in
oil. An encapsulated packet refers to a packet enclosed by a layer
of material. An encapsulated packet may refer to vesicle or other
microcapsule of liquid or gas that may contain a reagent, a sample,
a particle, a cell, an intermediate product, a final reaction
product, or any material. The surface of an encapsulated packet may
be coated with a reagent, a sample, a particle or cell, an
intermediate product, a final reaction product, or any material. An
example of an encapsulated packet is a lipid vesicle containing an
aqueous solution of reagent suspended in water. A solid packet
refers to a solid material that may contain, or be covered with a
reagent, a sample, a particle or cell, an intermediate product, a
final reaction product, or any material. An example of a solid
packet is a latex microsphere with reagent bound to its surface
suspended in an aqueous solution. Methods for producing packets as
defined herein are known in the art. Packets may be made to vary
greatly in size and shape, but in embodiments described herein,
packets may have a diameter between about 100 nm and about 1
cm.
[0013] In this respect, the invention includes a reaction surface,
an inlet port, means for generating a programmable manipulation
force upon the packet, a position sensor, and a controller. The
reaction surface is configured to provide an interaction site for
the packet. The inlet port is coupled to the reaction surface and
is configured to introduce the packet onto the reaction surface.
The means for generating a programmable manipulation force upon the
packet programmably moves the packet about the reaction surface
along arbitrarily chosen paths. As used herein, by "arbitrarily
chosen paths" it is meant that paths may be chosen to have any
shape about the reaction surface. Arbitrarily chosen paths are not
limited to movements that are predefined. Arbitrarily chosen paths
may be modified in an unlimited manner about the reaction surface
and may hence trace out any pattern. The position sensor is coupled
to the reaction surface and is configured to sense a position of
the packet on the reaction surface. The controller is coupled to
the means for generating a programmable manipulation force and to
the position sensor. The controller is configured to adjust the
programmable manipulation force according to the position.
[0014] In other aspects, the apparatus may also include an outlet
port coupled to the reaction surface. The outlet port may be
configured to collect the packet from the reaction surface. The
means for generating a manipulation force may include a conductor
adapted to generate an electric field. The means for generating a
manipulation force may include a light source. The manipulation
force may include a dielectrophoretic force, an electrophoretic
force, an optical force, a mechanical force, or any combination
thereof. The position sensor may include a conductor configured to
measure an electrical impedance of the packet. The position sensor
may include an optical system configured to monitor the position of
the packet. The means for generating a programmable manipulation
force and the position sensor may be integral.
[0015] In another respect, the invention is an apparatus for
microfluidic processing by programmably manipulating packets. The
apparatus includes a reaction surface, an inlet port, an array of
driving electrodes, and an array of impedance sensing electrodes.
As used herein, an "array" refers to any grouping or arrangement.
An array may be a linear arrangement of elements. It may also be a
two dimensional grouping having columns and rows. Columns and rows
need not be uniformly spaced or orthogonal. An array may also be
any three dimensional arrangement. The reaction surface is
configured to provide an interaction site for the packets. The
inlet port is coupled to the reaction surface and is configured to
introduce the packets onto the reaction surface. The array of
driving electrodes is coupled to the reaction surface and is
configured to generate a programmable manipulation force upon the
packets to direct the microfluidic processing by moving the packets
along arbitrarily chosen paths. The array of impedance sensing
electrodes is coupled to the reaction surface and is configured to
sense positions of the packets during the microfluidic
processing.
[0016] In other aspects, the apparatus may also include an outlet
port coupled to the reaction surface. The outlet port may be
configured to collect the packets from the reaction surface. The
apparatus may also include a controller coupled to the array of
driving electrodes and to the array of impedance sensing
electrodes. The controller may be adapted to provide a feedback
from the array of impedance sensing electrodes to the array of
driving electrodes. The array of driving electrodes and the array
of impedance sensing electrodes may be integral. The apparatus may
also include an integrated circuit coupled to the array of driving
electrodes and to the array of impedance sensing electrodes. The
apparatus may also include a coating modifying a hydrophobicity of
the reaction surface. The apparatus may also include a maintenance
port.
[0017] In another respect, the invention is an apparatus for
processing packets in a partitioning medium. As used herein, a
"partitioning medium" refers to matter that may be adapted to
suspend and compartmentalize other matter to form packets on a
reaction surface. A partitioning medium may act by utilizing
differences in hydrophobicity between a fluid and a packet. For
instance, hydrocarbon molecules may serve as a partitioning medium
for packets of aqueous solution because molecules of an aqueous
solution introduced into a suspending hydrocarbon fluid will
strongly tend to stay associated with one another. This phenomenon
is referred to as a hydrophobic effect, and it allows for
compartmentalization and easy transport of packets upon or over a
surface. A partitioning medium may also be a dielectric carrier
liquid which is immiscible with sample solutions. Other suitable
partitioning mediums include, but are not limited to, air, aqueous
solutions, organic solvents, oils, and hydrocarbons. The apparatus
includes a chamber, a programmable dielectrophoretic array, and an
impedance sensing array. As used herein, a "programmable
dielectrophoretic array" (PDA) refers to an electrode array whose
individual elements can be addressed with different electrical
signals. The addressing of electrode elements with electrical
signals may initiate different field distributions and generate
dielectrophoretic manipulation forces that trap, repel, transport,
or perform other manipulations upon packets on and above the
electrode plane. By programmably addressing electrode elements
within the array with electrical signals, electric field
distributions and manipulation forces acting upon packets may be
programmable so that packets may be manipulated along arbitrarily
chosen or predetermined paths. The chamber is configured to contain
the packets and the partitioning medium. The programmable
dielectrophoretic array is coupled to the chamber and is configured
to generate a programmable dielectrophoretic force to direct
processing of the packets. The impedance sensing array of
electrodes is integral with the programmable dielectrophoretic
array. The impedance sensing array of electrodes is configured to
sense a position of the packets within the chamber.
[0018] In other aspects, the apparatus may also include an
integrated circuit coupled to the programmable dielectrophoretic
array and to the impedance sensing array of electrodes. The
apparatus may also include a controller coupled to the programmable
dielectrophoretic array and to the impedance sensing array of
electrodes. The controller may be adapted to provide a feedback
from the impedance sensing array of electrodes to the programmable
dielectrophoretic array. The electrodes may be between about 1
micron and about 200 microns and may be spaced between about 1
micron and about 200 microns.
[0019] In another respect, the invention is a method for
manipulating a packet in which the following are provided: a
reaction surface, an inlet port coupled to the reaction surface,
means for generating a programmable manipulation force upon the
packet, a position sensor coupled to the reaction surface, and a
controller coupled to the means for generating a programmable
manipulation force and to the position sensor. A material is
introduced onto the reaction surface with the inlet port. The
material is compartmentalized to form the packet. A position of the
packet is sensed with the position sensor. A programmable
manipulation force is applied on the packet at the position with
the means for generating a programmable manipulation force. The
programmable manipulation force is adjustable according to the
position by the controller. The packet is programmably moved
according to the programmable manipulation force along arbitrarily
chosen paths.
[0020] In other aspects, the packet may include a fluid packet, an
encapsulated packet, or a solid packet. The compartmentalizing may
include suspending the material in a partitioning medium. The
material may be immiscible in the partitioning medium. The reaction
surface may include a coating, and the hydrophobicity of the
coating may be greater than a hydrophobicity of the partitioning
medium. The application of the programmable manipulation force may
include applying a driving signal to one or more driving electrodes
arranged in an array to generate the programmable manipulation
force. The programmable manipulation force may include a
dielectrophoretic force, an electrophoretic force, an optical
force, a mechanical force, or any combination thereof. The sensing
of a position may include applying a sensing signal to one or more
impedance sensing electrodes arranged in an array to detect an
impedance associated with the packet.
[0021] In another respect, the invention is a method of fluidic
processing in which the following are provided: a reaction surface,
an inlet port coupled to the reaction surface, an array of driving
electrodes coupled to the reaction surface, and an array of
impedance sensing electrodes coupled to the reaction surface. One
or more materials are introduced onto the reaction surface with the
inlet port. The one or more materials are compartmentalized to form
a plurality of packets. A sensing signal is applied to one or more
of the impedance sensing electrodes to determine a position of one
or more of the plurality of packets. A driving signal is applied to
one or more of the driving electrodes to generate a programmable
manipulation force on one or more of the plurality of packets at
the position. One or more of the plurality of packets are
interacted according to the programmable manipulation force.
[0022] In other aspects, at least one of the plurality of packets
may include a fluid packet, an encapsulated packet, or a solid
packet. The sensing signal and the driving signal may be a single
processing signal. The processing signal may include a first
frequency component corresponding to the sensing signal and a
second frequency component corresponding to the driving signal. A
packet distribution map may be formed according to the positions of
the plurality of packets. A position of one or more obstructions on
the reaction surface may be determined. The interacting of one or
more packets may include moving, fusing, merging, mixing, reacting,
metering, dividing, splitting, sensing, collecting, or any
combination thereof.
[0023] In another respect, the invention is a method for
manipulating one or more packets on a reaction surface in which the
following are provided: a programmable dielectrophoretic array
coupled to the reaction surface and an impedance sensing array of
electrodes integral with the programmable dielectrophoretic array.
A material is introduced onto the reaction surface. The material is
compartmentalized to form the one or more packets. A path is
specified upon the reaction surface. A programmable manipulation
force is applied with the programmable dielectrophoretic array on
the one or more packets to move the one or more packets along the
path. A position of the one or more packets is sensed with the
impedance sensing array of electrodes. Whether the position
corresponds to the path is monitored. The one or more packets are
interacted.
[0024] In other aspects, at lease one of the one or more packets
may include a fluid packet, an encapsulated packet, or a solid
packet. The method may also include sensing a position of an
obstruction; determining a modified path, the modified path
avoiding the obstruction; and applying a programmable manipulation
force on the one or more packets to move the one or more packets
along the modified path. The specification of a path may include
specifying an initial position and a final position. The
introduction of the material may include extracting the material
with a dielectrophoretic extraction force from an injector onto the
reaction surface. The interacting of one or more packets may
include moving, fusing, merging, mixing, reacting, metering,
dividing, splitting, sensing, collecting, or any combination
thereof.
[0025] Other features and advantages of the present invention will
become apparent with reference to the following description of
typical embodiments in connection with the accompanying drawings
wherein like reference numerals have been applied to like elements,
in which:
BRIEF DESCRIPTION OF THE DRAWINGS
[0026] FIG. 1 is a simplified schematic diagram that illustrates a
microfluidic device according to one embodiment of the presently
disclosed method and apparatus.
[0027] FIG. 2 is a simplified illustration of dielectrophoretic
force phenomenon.
[0028] FIG. 3 illustrates a position sensing system according to
one embodiment of the presently disclosed method and apparatus.
[0029] FIG. 4 is a three dimensional view of a microfluidic device
according to one embodiment of the presently disclosed method and
apparatus.
[0030] FIG. 5 is a side cross sectional view of a microfluidic
device according to one embodiment of the presently disclosed
method and apparatus.
[0031] FIG. 6 is a simplified block representation of a
microfluidic system according to one embodiment of the presently
disclosed method and apparatus.
[0032] FIG. 7 is a simplified block representation of a signal
application arrangement according to one embodiment of the
presently disclosed method and apparatus.
[0033] FIG. 8 is a cross sectional view of microfluidic device
according to one embodiment of the presently disclosed method and
apparatus.
[0034] FIG. 9 is a top view of a microfluidic device according to
one embodiment of the presently disclosed method and apparatus.
[0035] FIG. 9B is another top view of a microfluidic device
according to one embodiment of the presently disclosed method and
apparatus.
[0036] FIG. 10 is a simplified block representation of a
microfluidic system according to one embodiment of the presently
disclosed method and apparatus.
[0037] FIG. 11 is a top view of a microfluidic device showing a
microfluidic process according to one embodiment of the presently
disclosed method and apparatus.
[0038] FIG. 12 illustrates certain packet interactions according to
one embodiment of the presently disclosed method and apparatus.
[0039] FIG. 13 is a flow chart showing a microfluidic process
according to one embodiment of the presently disclosed method and
apparatus.
DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0040] The disclosed method and apparatus provide many advantages.
For instance, they permit the fluidic processing of minute
quantities of samples and reagents. The apparatus need not use
conventional hardware components such as valves, mixers, pump. The
apparatus may be readily miniaturized and its processes may be
automated or programmed. The apparatus may be used for many
different types of microfluidic processing and protocols, and it
may be operated in parallel mode whereby multiple fluidic
processing tasks and reactions are performed simultaneously within
a single chamber. Because it need not rely on narrow tubes or
channels, blockages may be minimized or eliminated. Further, if
obstructions do exist, those obstructions may be located and
avoided with position sensing techniques.
[0041] Allowing for flexible microfluidic processing, the disclosed
method and apparatus has vast applications including, but not
limited to, blood and urine assays, pathogen detection, pollution
monitoring, water monitoring, fertilizer analysis, the detection of
chemical and biological warfare agents, food pathogen detection,
quality control and blending, massively parallel molecular
biological protocols, genetic engineering, oncogene detection, and
pharmaceutical development and testing.
[0042] In one embodiment of the disclosed method and apparatus, a
fluidic device 10 as shown in FIG. 1 is employed. As illustrated,
fluidic device 10 may include a reaction surface 12, a port 15,
packets 21, wall 22, position sensor 23, a force generator 25, and
a controller 81.
[0043] In operation, one or more materials may be introduced onto
reaction surface 12 through port 15. The one or more materials may
be compartmentalized to form packets 21 within a partitioning
medium (not shown). Force generator 25 generates a manipulation
force on packets 21 to facilitate fluidic manipulations and
interactions. In the illustrated embodiment, force generator 25
generates two forces, F.sub.1 and F.sub.2, that manipulate packets
21 and moves them according to the dashed lines of FIG. 1. Position
sensor 23 senses the positions of packets 21 and is able to monitor
any packet interactions. As position sensor 23 is coupled to force
generator 25 by controller 81, a feedback relationship may be
established. Such feedback may include determination of the
position of packets 21 on reaction surface 12 that allows for the
application of manipulation forces on packets 21 based on position
information. The position of packets during manipulation may thus
be continuously monitored and this information may be used to
continuously adjust one or more manipulation forces so to achieve
movement of packets 21 along a desired trajectory to a desired
location on reaction surface 12.
[0044] In the illustrated embodiment of FIG. 1, forces F.sub.1 or
F.sub.2 may include many different types of forces. For instance,
forces F.sub.1 and F.sub.2 may be dielectrophoretic,
electrophoretic, optical (as may arise, for example, through the
use of optical tweezers), mechanical (as may arise, for example,
from elastic traveling waves or from acoustic waves), or any other
suitable type of force (or combination thereof). In one embodiment,
forces F.sub.1 and F.sub.2 may be programmable. Using programmable
forces, packets may be manipulated along arbitrarily chosen
paths.
[0045] In the illustrated embodiment of FIG. 1, position sensor 23
may be operated with various mechanisms to sense positions of
packets 21. For instance, an optical imaging system may be used to
determine and monitor packet positions. Specifically, an optical
microscope may be connected to a CCD imaging camera, which may be
interfaced with an imaging card in a computer. The information from
the imaging card may be processed in the computer using
image-analysis software. Alternatively, a CCD imaging device may be
incorporated in or above the reaction surface 12 to monitor the
positions of packets. Thus, positions of packets and their movement
on reaction surface 12 may be continuously monitored and recorded
in the computer. A different mechanism of packet position sensing
uses electrical impedance measurements. The presence or absence of
a packet between two electrode elements may affect the electrical
impedance between the electrodes. Thus, measurement of electrical
impedance between electrode elements may allow for indirect
monitoring of packet positions.
[0046] In order to better understand the operation and design of
the currently disclosed method and apparatus, which will be
discussed first in relation to dielectrophoretic forces, it is
useful to discuss dielectrophoretic theory in some detail. Such a
discussion is aided by FIG. 2, which illustrates two packets, 21a
and 21b, both being subjected to dielectrophoretic forces.
[0047] Dielectrophoretic forces may arise when a packet is placed
in an inhomogeneous electrical field (AC or DC). In FIG. 2 the
electrical field is weaker on the left side than on the right side.
An electrical field induces electrical polarizations in the packet.
The polarization charges are depicted at the two ends of the
packets 21a and 21b along the field lines 35. Dielectrophoretic
forces result from the interaction between the induced polarization
(labeled as m.sub.1 and m.sub.2 in FIG. 2) and the applied
inhomogeneous field. If a packet is suspended in a medium having
different dielectric properties, such as a partitioning medium,
then the packet may remain compartmentalized and may readily
respond to manipulation forces against viscous drag. In a field of
non-uniform strength, a packet may be directed towards either
strong (packet 21a) or weak (packet 21b) electrical field regions,
depending on whether the packet is more (packet 21a) or less
(packet 21b) polarizable than a partitioning medium. In a field of
non-uniform phase distribution (i.e. a traveling electrical field),
a packet may be directed towards field regions of larger or smaller
phase distribution, depending whether the packet has a longer or
shorter dielectric response time than that of a partitioning
medium.
DEP Theory
[0048] When a packet of radius r, suspended in an immiscible medium
of different dielectric properties, is subjected to an electrical
field of frequency f, the polarization of the packet can be
represented using an effective dipole moment (Wang et al., "A
Unified Theory of Dielectrophoresis and Traveling Wave
Dielectrophoresis", Journal of Physics D: Applied Physics, Vol 27,
pp. 1571-1574, 1994, incorporated herein by reference)
{right arrow over (m)}(f)=4.pi..epsilon..sub.m
r.sup.3P.sub.CM(f){right arrow over (E)}(f) (1)
where {right arrow over (m)}(f) and {right arrow over (E)}(f) are
the dipole moment and field vectors in the frequency domain,
P.sub.CM(f) is the so-called Clausius-Mossotti factor, given by
P.sub.CM(f)=(.epsilon.*.sub.d-.epsilon.*.sub.m)/(.epsilon.*.sub.d+2.epsi-
lon.*.sub.m). (2)
[0049] Here
.epsilon.*.sub.k=.epsilon..sub.k-j.sigma..sub.k/(2.pi.f) are the
complex permittivities of the packet material (k=d) and its
suspension medium (k=m), and .epsilon. and .sigma. refer to the
dielectric permittivity and electrical conductivity, respectively.
Using the effective dipole moment method, the DEP forces acting on
the packet are given by
{right arrow over
(F)}(f)=2.pi.r.sup.3.epsilon..sub.m(Re[P(f)].gradient.E.sub.(rms).sup.2+I-
m[P(f)](E.sub.x0.sup.2.gradient..phi..sub.x0+E.sub.y0.sup.2.gradient..phi.-
.sub.y0+E.sub.z0.sup.2.gradient..phi..sub.z0)) (3)
where E(rms) is the RMS value of the field strength, E.sub.i0 and
.phi..sub.i0 (i=x; y;z) are the magnitude and phase, respectively,
of the field components in a Cartesian coordinate frame. Equation
(3) shows that the DEP force contains two independent terms. The
first, relating to the real (in phase) part of the polarization
factor Re[P(f)] and to non-uniformities in the field magnitude
(.gradient.E.sub.(rms).sup.2). Depending on the sign of Re[P(f)],
this force directs the packet either toward strong or weak field
regions. The second term relates to the imaginary (out of phase)
part of the polarization factor (Im[P(f)]) and to field phase
non-uniformities (V.phi..sub.i0, i=x; y; z) that correspond to the
field traveling through space from large to small phase regions.
Depending on the sign of Im[P(f)], this directs packets toward
regions where the phase values of the field components are larger
or smaller.
[0050] Equations (1-3) indicate that the DEP phenomena have the
following characteristics:
[0051] (1) DEP forces experienced by packets are dependent on the
dielectric properties of the packets (.epsilon.*.sub.d) and the
partitioning medium (.epsilon.*.sub.m).
[0052] (2) The strong dependence of three-dimensional DEP forces on
the field configuration allows for versatility in implementing
dielectrophoretic manipulations.
DEP Forces on Packets
[0053] In one embodiment, a conventional dielectrophoresis
component may be used for packet manipulation. In this case, the
DEP force is given by
{right arrow over (F)}(f)=2.pi.r.sup.3.epsilon..sub.m
Re[P(f)].gradient.E.sub.(rms).sup.2 (4)
[0054] where r is the packet radius, .epsilon..sub.m is the
dielectric permittivity of the suspending fluid. Re[P(f)] is the
real (in phase) part of the polarization factor and
.gradient.E.sub.(rms).sup.2 is the is field non-uniformity factor.
For packets of water (.epsilon.=78 and .sigma.>10.sup.-4 S/m)
suspended in a hydrocarbon fluid (.epsilon.=.about.2 and
.sigma..about.0), the factor Re[P(f)] is always positive and close
to unity. Therefore, water packets are always attracted towards
regions of large field strength. For example, if an electrode array
composed of circular electrodes arranged in a hexagonal fashion is
provided, water packets may be dielectrophoretically moved towards
and trapped between, for example, an electrode pair, over a single
electrode, or above a plurality of electrodes to which electrical
signals are applied. Switching the electrical signals may result in
movement of the DEP traps and may cause water packets to move in a
chamber. Thus, packet manipulation may be realized by switching
electrical signals applied to an electrode array so that DEP field
traps are made "mobile" within a chamber.
Typical Forces and Velocities
[0055] For a water packet of 100 .mu.m suspended in a hydrocarbon
fluid such as decane, the DEP force may be on the order of 1000 pN
if the field non-uniformity is 1.25.times.10.sup.13 V.sup.2/m.sup.3
(equivalent to 5V RMS applied to an electrode pair of distance 50
.mu.m with the field decaying to zero at 1000 .mu.m). If the
viscosity of the hydrocarbon fluid is small (0.838 mPa for Decane),
the packet velocity may be of the order of 600 .mu.m/sec,
indicating that fast manipulation of packets is possible with
electrode arrays. In the above analysis, DEP force equation (4) has
been used, which was developed for non-deformable particles and
holds well for suspended particles (such as cells, latex to
particles). Fluid packets may be deformed under the influence of
applied electrical field, ii affecting the accuracy of equation (4)
in describing DEP forces for packets. Nevertheless, equation (4)
should be generally applicable with some possible correction
factors for different packet shapes.
[0056] FIG. 3 shows one possible implementation of position sensor
23 of FIG. 2. is Shown in FIG. 3 are five impedance sensing
electrodes 19, here illustrated as 19a, 19b, 19c, 19d, and 19e.
Each sensing electrode 19 may be coupled to an impedance sensor 29,
here illustrated as impedance sensors 29a, 29b, 29c, and 29d. In
one embodiment, impedance sensing electrodes 19 may be positioned
in operative relationship with surface 12 of fluidic device 10 in
FIG. 1. For instance, sensing electrodes 19 may be placed on or
near surface 12. As packets 21 are manipulated about surface 12 by
the application of appropriate manipulation forces, impedance
sensing electrodes 19 and sensors 29 may sense a position of
packets 21 by making one or more impedance measurements.
[0057] If the dielectric medium above an electrode is displaced by
a packet having different dielectric and/or conductive properties,
the impedance detected at the electrode element will change. Thus,
one may determine the position of packets 21 by noting the
impedance measurements associated therewith. As is shown in FIG. 3,
the impedance between impedance sensing electrodes 19a and 19b is
"high" (see impedance sensor 29d) relative to, for instance, the
impedance between impedance sensing electrodes 19b and 19c (see
impedance sensor 29c). Thus, by pre-determining that the "high"
impedance value corresponds to the impedance due to the
partitioning medium, it may be deduced that some material of
different impedance to the partitioning medium lies somewhere
between impedance sensing electrodes 19d and 19e and between 19b
and 19c because the impedance associated with those electrodes is
"low" (see impedance sensor 29a). By like reasoning, one may assume
that no packet lies between impedance sensing electrodes 19c and
19d, for the impedance between those two electrodes is relatively
"high" (see impedance sensor 29b and 29c).
[0058] Those of skill in the art will appreciate that the "low" and
"high" values discussed above may be reversed, depending upon the
relative impedances of a packet and of a suspending medium. In
other words, in some situations, a relatively "high" impedance
measurement may signal the presence of a packet in between a pair
of electrodes while a relatively "low" impedance may signal the
lack of a packet. Those of skill in the art will also appreciate
that individual impedance measurements may exhibit a wide range of
values (not just "low" or "high"), and it may be possible to
characterize different types and sizes of materials by noting their
associated impedance measurements. For instance, one may be able to
differentiate, by type, the two packets 21 of FIG. 3 by noting any
differences in their impedance readings on impedance sensors 29a
and 29c.
[0059] Impedance sensing may be based on the so-called mixture
theory, which associates the impedance of a heterogeneous system
with the dielectric properties of various system components and
their volume fractions. Take a two-component, heterogeneous system
where component 2 having complex dielectric permittivity
( 2 * = 2 - j .sigma. 2 2 .pi. f , ##EQU00001##
f is the frequency) and a volume fraction .alpha. is suspended in
component 1 having complex dielectric permittivity
( 1 * = 1 - j .sigma. 1 2 .pi. f ) . ##EQU00002##
The complex permittivity of the total system is given by (Wang et
al., "Theoretical and experimental investigations of the
interdependence of the dielectric, dielectrophoretic and
electrorotational behavior of colloidal particles" in J. Phys. D:
Appl. Phys. 26: 312-322, 1993, incorporated herein by
reference)
sys * = 1 * 1 .alpha. + 2 2 * - 1 * 2 * + 1 * 1 .alpha. - 2 * - 1 *
2 * + 1 * . ##EQU00003##
[0060] The total impedance of the system, which is assumed to have
length L and cross-sectional area A, is given by
.OMEGA. = L .omega. sys * A . ##EQU00004##
[0061] The electrical impedance between two electrode elements in
the presence or absence of a packet may be analyzed using the above
equations, with the parameters L and A determined experimentally.
The existence of a packet may correspond to .alpha.>0 and the
absence of a packet may correspond to .alpha.=0. From these
equations, an impedance change would occur when a packet having
different dielectric property (.epsilon.*.sub.2) from the
partitioning media (.epsilon.*.sub.1) is introduced into the space
between the two electrode elements.
[0062] A relatively low impedance measurement may indicate an
obstruction or a packet (as is illustrated in FIG. 3) on or near
surface 12. By determining impedance values, one may map locations
of obstructions or packets relative to surface 12. In this way, one
may generate a packet and/or obstruction distribution map with
respect to reaction surface 12 of fluidic device 10. With the
benefit of this disclosure, one of skill in the art will appreciate
that the description associated with FIG. 3 may be implemented in
many different ways. In particular, one may use any suitable type
of impedance measurement devices known in the art to function with
one or more electrodes. Such devices may include an impedance
analyzer, a DC/AC conductance meter, or any circuit based upon
methods of operation of these or other instruments having similar
function.
[0063] FIG. 4 shows a three dimensional view of one embodiment of a
fluidic device 10 according to the present disclosure. Fluidic
device 10 includes reaction surface 12, an inlet port 14, an outlet
port 16, driving electrodes 18, impedance sensing electrodes 19,
connectors 20, and wall 22.
[0064] Reaction surface 12 provides an interaction site for
packets. In one embodiment, reaction surface 12 may be completely
or partially covered with a partitioning medium (not shown in FIG.
4) or other substance. In one embodiment, reaction surface 12 may
be coated. In particular, for manipulation of aqueous packets in a
hydrophobic partitioning medium, reaction surface 12 may include a
hydrophobic coating, or layer, having a hydrophobicity similar to
or greater than the hydrophobicity of the partitioning medium. Such
a coating may prevent an aqueous packet from sticking, from
spreading, or from becoming unstable upon contact with reaction
surface 12. Additionally, a coating may modify association and/or
interaction forces between packets and reaction surfaces to
facilitate manipulation of packets by appropriate manipulation
forces. Further, a coating may be used to reduce contamination of
reaction surfaces by reagents in packets. Still further, a coating
may facilitate the deliberate adhesion, wetting, or sensing of
packets at or on reaction surfaces. If a dielectric layer coating
is applied, the layer should be made sufficiently thin to allow AC
electric field penetration through the dielectric layer. In one
embodiment, the thickness of the layer may be between about 2 nm
and about 1 micron. In one embodiment, a hydrophobic coating may be
Teflon that may be applied by means known in the art such as
sputtering or spin-coating. It is to be understood that any other
suitable coating that modifies an interaction between packets and
the reaction surface may be used.
[0065] Reaction surface 12 may be formed from a number of suitable
materials. In the illustrated embodiment, reaction surface 12 is a
planar surface that has an upper surface including driving
electrodes 18 and impedance sensing electrodes 19. Although
illustrated as being coplanar with reaction surface 12, it is to be
understood that driving electrodes 18 and 19 may also be elevated
or depressed with respect to reaction surface 12. Likewise,
reaction surface 12 need not be planar. Rather, it may have concave
or convex portions, or it may be deformed in some other manner.
Reaction surface 12 may be glass, silicon dioxide, a polymer, a
ceramic, or any suitable electrically insulating material. The
dimensions of reaction surface 12 may vary widely depending on the
application but may be between about 20 microns by about 20 microns
and about 50 centimeters by about 50 centimeters. More
particularly, reaction surface 12 may be between about 3
millimeters by about 3 millimeters and about 30 centimeters by
about 30 centimeters.
[0066] Inlet port 14 may be adapted to inject or introduce
materials onto reaction surface 12 and may be any structure
allowing ingress to reaction surface 12. In the illustrated
embodiment, inlet port 14 consists of an opening in wall 22. Such
an opening may be of any suitable size or shape. Alternatively,
inlet port 14 may be a syringe needle a micropipette, a tube, an
inkjet injector, or any other suitable device able to inject a ii
material for introduction onto reaction surface 12. Using a
micropipette or equivalent device, wall 22 may not need to include
any openings. Rather, material may be introduced onto reaction
surface 12 from above. A micropipette or any other equivalent
device may be attached to a micromanipulation stage (not shown in
FIG. 4) so that is material may be precisely deposited onto
specific locations of reaction surface 12. In one embodiment, inlet
port 14 may consist of a cylindrical tube opening onto reaction
surface 12. Such a tube may have a diameter of between about 1
micrometer and about 1 mm and, more particularly, between about 10
and 100 microns.
[0067] Outlet port 16 may be adapted to collect packets of material
from reaction surface 12. Outlet port 16 may be any structure
allowing egress from reaction surface 12. In the illustrated
embodiment, outlet port 16 consists of an opening in wall 22. The
opening may be of any suitable size or shape. Alternatively, outlet
port 16 may be a micropipette or any other equivalent device able
to collect a material from reaction surface 12. Wall 22 may not
need to include any openings. Rather, packets of material may be
collected from reaction surface 12 from above. A syringe or any
other equivalent device may be attached to a micromanipulation
stage (not shown in FIG. 4) so that packets may be precisely
collected from specific locations on reaction surface 12. In one
embodiment, outlet port 16 may consist of a cylindrical tube
opening onto reaction surface 12. Such a tube may have a diameter
of about 1 millimeter and a length of about 3 centimeters or
longer.
[0068] In one embodiment, inlet port 14 and outlet port 16 may be
integral. For instance, in the embodiment shown in FIG. 1 port 15
is a cylindrical tube opening onto reaction surface 12. In
alternative embodiments, one micropipette may serve as both an
inlet port and an outlet port. Alternatively, a single opening in
wall 22 may serve both input and output functions. In another
embodiment, multiple inlet and outlet ports may be utilized.
[0069] Fluidic device 10 may include an arbitrary number of inlet
and outlet ports. For example, any one of the three unnumbered
openings in wall 22, illustrated in FIG. 4, may serve as an inlet
port, an outlet port, or an integral inlet-outlet port, such as
port 15 of FIG. 1. In another embodiment, multiple inlet and/or
outlet ports may extend completely or partially along a wall 22 so
that materials may be introduced and/or collected to and/or from
reaction surface 12. In such an embodiment, one may more precisely
introduce or collect materials.
[0070] In FIG. 4, driving electrode 18 is one of a number of other
driving electrodes arranged in an array upon reaction surface 12.
In this embodiment, driving electrodes 18 may be associated with
force generator 25 of FIG. 1, for the driving electrodes 18 may
contribute to the generation of forces, such as forces F.sub.1 and
F.sub.2 of FIG. 1, to manipulate packets of material on reaction
surface 12 to promote, for instance, microfluidic interactions.
[0071] Dielectrophoretic forces may be generated by an array of
individual driving electrodes 18 fabricated on an upper surface of
a reaction surface 12. The driving electrode elements 18 may be
individually addressable with AC or DC electrical signals. Applying
an appropriate signal to driving electrode 18 sets up an electrical
field that generates a dielectrophoretic force that acts upon a
packet, known to be at a certain location through impedance
measurements as described above in relation to FIG. 3. Switching
different signals to different electrodes sets up electrical field
distributions within fluidic device 10. Such electrical field
distributions may be utilized to manipulate packets in a
partitioning medium.
[0072] In particular, the movement of packets under the influence
of a manipulation force may be controlled by switching appropriate
electrical signals to different combinations of driving electrodes
18. Specifically, the switching of electrical signals may initiate
different field distributions and generate manipulation forces that
trap, repel, transport, or perform other manipulations upon packets
of material. By programmably switching electrical signals to
different combinations of driving electrodes 18 within an array,
electric field distributions and manipulation forces acting upon
packets may be programmable so that packets may be manipulated
along arbitrarily chosen or predetermined paths in a partitioning
medium along reaction surface 12. Thus, packets may be manipulated
in an unlimited manner. Signals may be appropriately switched to
cause, for instance, a packet to move a single "unit distance"--a
distance between two neighboring electrodes. Further, by
programmably switching electrical signals, different microfluidic
reactions may be performed in series or in parallel. An electrode
array having such an ability to utilize programmable
dielectrophoretic forces by programmed switching of electrical
signals to different combinations of driving electrodes 18 may be
termed a programmable dielectrophoretic array (PDA).
[0073] In FIG. 4, impedance sensing electrode 19 is one of a number
of other impedance sensing electrodes arranged in an array upon
reaction surface 12. In this embodiment, impedance sensing
electrodes 19 may be associated with position sensor 23 of FIG. 1
and is illustrated in FIG. 3. Impedance sensing electrodes 19
contribute to the sensing of packet positions upon reaction surface
12 so that those packets of material may be monitored and
manipulated according to position.
[0074] In the illustrated embodiment, driving electrodes 18 and
impedance sensing electrodes 19 are electrodes of a two dimensional
electrode array coupled to a top surface of reaction surface 12.
The size of the array may vary according to need, but in one
embodiment a 16 by 16 array is employed. Because fluidic device 10
is scaleable, smaller or larger arrays may be fabricated without
significant departure from the present disclosure. For example, 256
by 256 arrays or larger may be made according to the present
disclosure. Driving electrodes 18 and impedance sensing electrodes
19 within an array may be uniformly or non-uniformly spaced. The
spacing may vary widely, but in one embodiment, the spacing may be
between about 2 microns and about 200 microns. The electrodes may
have different forms such as lines, squares, circles, diamonds,
polygons, or other suitable shapes. The dimensions of each
electrode may vary, but a typical electrode may be between about
0.2 microns and about 10 mm., and more particularly, between about
1 micron and about 200 microns. Driving electrodes 18 and impedance
sensing electrodes 19 may be formed using any method known in the
art. In one embodiment, such electrodes may be formed using
standard photolithography techniques. For example, one may refer
to, e.g., D. Qin et al, "Microfabrication, Microstructures and
Microsystems", Microsystem Technology in Chemistry and Life
Sciences (Ed. Manz and Becker), Springer, Berlin, 1997, pp 1-20,
which is incorporated herein by reference. Also, one may refer to
Madou, Fundamentals of Microfabrication, CRC Press, Boca Raton,
1997, which is incorporated herein by reference. Depending upon the
particular application, and the nature of the packets and
partitioning medium, the size and spacing of electrodes 18 and 19
may be smaller than, of similar size, or larger than the diameters
of the packets.
[0075] In one embodiment, impedance sensing electrodes 19 may be
integral with driving electrodes 18. In such an embodiment, the
resulting array may be termed an integral array. With an integral
array, a single conductor coupled to reaction surface 12 may serve
both purposes--driving packets and sensing positions of packets.
Thus, a programmable manipulation force may be generated upon
packets upon reaction surface 12 and a position of those packets
may be sensed with a single electrode array.
[0076] In the embodiment of FIG. 4, wall 22 is adapted to enclose
one or more sides of reaction surface 12. It is to be understood
that wall 22 may be any suitable structure capable of enclosing one
or more sides and/or the top of reaction surface 12. As
illustrated, wall 22 encloses four sides of reaction surface 12,
defining an open reaction surface chamber. In a most typical
embodiment, the chamber may have a thickness of between about 10
microns and about 20 millimeters. In another embodiment, wall 22
may enclose the top of reaction surface 12, forming a closed
reaction chamber.
[0077] Wall 22 may be formed from any suitable material. In one
embodiment, wall 22 may be made from machined plastic, aluminum,
glass, plastic, ceramic, or any combination thereof. In one
embodiment, wall 22 may be partially or completely transparent to
certain wavelengths of radiation. Thus, radiation may be
transmitted through wall 22 to initiate or maintain certain
microfluidic reactions or processes for sensing. For instance, a
photochemical reaction may be initiated through wall 22.
[0078] Connectors 20 of FIG. 4 may be adapted to provide electrical
connections to driving electrodes 18 and to impedance sensing
electrodes 19. Connectors 20 may provide electrical connections to
an entire array of electrodes, or to preselected ones or groups. In
one embodiment, connectors 20 are coupled to a controller (not
shown in FIG. 4) that may adjust a programmable manipulation force
distribution generated by driving electrodes 18 according one or
more packets position sensed with impedance sensing electrodes 19.
Thus, such a controller may effectively provide a feedback
mechanism between the driving electrodes 18 and the impedance
sensing electrodes 19--The signals applied to driving electrodes 18
may be adjusted in view of one or more results from the impedance
sensing electrodes 19.
[0079] Turning now to FIG. 5, there is shown a side cross section
view of a fluidic device 10 according to the present disclosure.
Fluidic device 10 includes a reaction chamber 41 and an array of
integral impedance sensing and driving electrodes, integral array
43. In the illustrated embodiment, a control chip 60 is coupled to
integral array 43. Positioned upon a top surface of control chip 60
may be capillary wall 62 that forms a lower surface of a capillary
64. Capillary 64 may lead to an inlet port 14 that leads into
chamber 41. Although illustrated with only one inlet port, it is
contemplated that there may be several such ports providing access
to chamber 41. Above capillary 64 is a substrate 66 that, in one
embodiment, is made of glass although any suitable material known
in the art may be utilized instead.
[0080] In one embodiment, control chip 60 may be an integrated
circuit configured to control integrated array 43. Alternatively,
control chip 60 may be a control interface leading to another
controlling device such as an integrated circuit, computer, or
similar device that may control integral array 43. Control chip 60
may utilize flip-chip technology or any other suitable technique to
establish electrical control over integral array 43 by switching
different signals to and from it.
[0081] FIG. 6 shows a controller 81 according to one embodiment of
the presently disclosed method and apparatus. Controller 81 may
include a computer 80, a signal generator 82, an electrode selector
84, a transducer 88, and a fluidic device 10 having a driving
electrode 18 and an impedance sensing electrode 19.
[0082] Computer 80 may be configured to control fluidic device 10
and the fluid processing occurring upon reaction surface 12.
Computer 80 may have a user interface that allows for simple
programming of signal generator 82 and transducer 88, which
measures impedance, to allow for programmable microfluidic
processing. In particular, computer 80 may programmably control the
initiation/termination of one or more signals from signal generator
82, the parameters of the one or more signals including
frequencies, voltages, and particular waveforms, and control the
switching of one or more signals from generator 82 to different
combinations of electrodes 18 and 19.
[0083] Computer 80 may vary signals in many ways. For instance, one
signal having a first frequency component may be sent through
electrode selector 84 to a driving electrode 18 while another
signal having a second, different frequency component may be sent
to, for instance, an impedance sensing electrode 19 and through
electrode selector 84. Any sequence of signals or combinations of
signals may be sent different combinations of electrodes and from
the fluidic device 10. Any signal parameter may be varied and any
electrode selection may be controlled so that appropriate electric
fields may be established at particular locations upon reaction
surface 12. Alternating Current or Direct Current signals may be
utilized.
[0084] Signal generator 82 may send a driving signal to one or more
driving electrodes 18 while sending a sensing signal to one or more
impedance sensing electrodes 19. In one embodiment, the driving
signal and the sensing signal may comprise a single, composite
processing signal having different frequency components. Such a
signal may be used with an integrated array to provide (via a
single processing signal) a frequency component to generate a
programmable manipulation force and a frequency component to
provide an impedance sensing signal. The manipulation and impedance
sensing components may also be combined by multiplexing or
switching in time as is known in the art.
[0085] In one embodiment, signal generator 82 provides one or more
programmable driving signals to one or more driving electrodes 18
through electrode selector 84 so that a programmable
alternating-current electric field, such as a non-uniform field,
may be produced at reaction surface 12. That electric field may
induce polarization of packets of materials adjacent to or in the
vicinity of the one or more driving electrodes 18. A programmable
dielectrophoretic force distribution may, in this manner, be
generated that manipulates packets in a controllable, programmable
manner so that varied programmable fluidic interactions may take
place upon reaction surface 12.
[0086] In one embodiment, signal generator 82 provides a sensing
signal to one or more impedance sensing electrodes 19 so that an
impedance measurement may be made. The impedance sensing signal may
be applied to one or more pairs of impedance sensing electrodes 19
and a change in voltage or current may be detected and transmitted
to computer 80 via sensing electrodes 88 and wire 86. Computer 80
may then compute the impedance and hence, determine whether a
packet or obstruction was present at or near the pair(s) of
impedance sensing electrodes 19 being probed.
[0087] In an embodiment utilizing a single integrated array
(instead of separate impedance sensing and driving electrode
arrays, an integrated array utilizes electrodes that function to
both drive and sense packets), the integrated array may both
generate a programmable manipulation force and sense an impedance.
In one approach, electrical sensing signals for sensing electrode
impedance may be applied at different frequencies from driving
signals for manipulation of packets. Summing signal amplifiers (not
shown) may be used to combine signals from sensing and driving
electronics. By using a frequency filter network (not shown),
electrode impedance sensing signals may be isolated from the
driving signals. For example, a constant current at sensing
frequency f.sub.s may be applied to integrated electrode pairs to
be measured. The sensing electronics 88, may then be operated at
only the applied frequency f.sub.s to determine a voltage drops
across the integrated electrode pairs, thus allowing the impedance
at the sensing frequency f.sub.s to be derived without interference
from the driving signals.
[0088] In another embodiment, driving signals may be used to
monitor electrical impedance directly. Driving signals may be
switched to one or more integrated electrodes to generate a force
to manipulate or interact packets upon a reaction surface.
Simultaneously, an electrical current sensing circuit may be used
to measure electrical current going through the energized
integrated electrodes. Electrode impedances may be derived from
such measurements of electrical current.
[0089] Although any suitable device may be used, in one embodiment
a function generator is used as signal generator 82. More
particularly, an arbitrary waveform signal generator in combination
with voltage or power amplifies or a transformer may be used to
generate the required voltages. In one embodiment, signal generator
82 may provide sine-wave signals having a frequency up to the range
of GHz and more particularly between about 1 kHz and about 10 MHz
and a voltage between about 1 V peak-to-peak and about 1000 V
peak-to-peak, and more particularly, between about 10 V
peak-to-peak and about 100 V peak-to-peak.
[0090] As illustrated, signal generator 82 may be connected to an
electrode selector 84. Electrode selector 84 may apply one or more
signals from signal generator 82 to one or more individual
electrodes (impedance sensing electrodes and/or driving electrodes
may be individually addressable). Electrode selector 84 may be one
of a number of suitable devices including a switch, a multiplexer,
or the like. Alternatively, electrode selector 84 may apply one or
more signals to one or more groups of electrodes. In one
embodiment, selector 84 is made of electronic switches or a
multiplexer. Selector 84 may be digitally controlled. With the
benefit of this disclosure, those of skill in the art will
understand that selector 84 may be any suitable device that may
programmably divert one or more signals to one or more electrodes
in any arbitrary manner.
[0091] As illustrated in FIG. 6, controller 81 provides a feedback
loop mechanism from impedance sensing electrodes 19 to driving
electrodes 18 via computer 80, which itself is coupled to signal
generator 82, selector 84, and transducer 88. With the benefit of
the present disclosure, those of skill in the art will recognize
that controller 81 may contain more or fewer components. The
feedback mechanism allows computer 80 to tailor its commands to
signal generator 82 according to positions of packets upon reaction
surface 12, as determined by impedance sensing electrodes 19. Thus,
controller 81 allows for the adjustment of driving signals (and
hence the adjustment of programmable manipulation forces) according
to positions of packets (as determined by impedance sensing
electrodes 19). In embodiments utilizing an integral array of
electrodes having integral impedance sensing electrodes 19 and
driving electrodes 18, a feedback mechanism may operate as is
follows. Positions of packets may be determined by measuring
impedances between electrical elements by applying impedance
sensing signals to the integral array. Position information may
then be used to control driving signals to the integral array to
perform microfluidic processing through the manipulation of
packets. In one embodiment computer 80 may be replaced by an
application specific integrated circuit controller (ASIC) designed
specifically for the purpose.
[0092] FIG. 7 shows an electrode driver 94 according to an
embodiment of the presently disclosed method and apparatus. Driver
94 includes a computer 80, a signal generator 82, a resistor
network 100, a switching network 104, and a bitmap 108. Driver 94
is coupled to fluidic device 10 which includes reaction surface 12
and an integral array 43.
[0093] Driver 94 may assist in the application of signals to
integral array 43 in order to direct microfluidic interactions of
packets of material upon reaction surface 12. In one embodiment,
computer 80 directs signal generator 82 to apply an AC signal to
integral array 43. In the illustrated embodiment, from signal
generator 82 there may be provided, for example, eight increasing
voltage amplitudes using resistor network 100, although more or
fewer voltage amplitudes may be used. The eight AC signals may be
distributed by switching network 104 via connection 106 to the
integral array 43 according to a bitmap 108 or according to any
other suitable data structure stored in computer 80 or in another
device. By modifying bitmap 108 via computer 80, different voltage
amplitudes may be applied to different electrodes.
[0094] In one embodiment, signals to each electrode of integral
array 43 may be represented in bitmap 108 by 3 bits to address
eight available voltage amplitudes. Voltage amplitude distributions
of bitmap 108 may be transmitted sequentially to switching network
104 via connection 110 twelve bits at a time using a communication
io protocol as is known in the art. In one embodiment, the
communication protocol may use the following convention. To address
a single electrode of integral array 43, the first four bits may
specify the row of the array. The second four bits may specify the
column of the array. The next three bits may specify the desired
voltage to be applied. The last bit may be used for error control
by parity check. The rows/column arrangement may be used for
different layouts of arrays. For instance, the row/column
convention of addressing may be used even for a hexagonal grid
array configuration. Those skilled in the art will appreciate that
other methods may be used to address the electronic switching
network 104 from computer 80.
[0095] FIG. 8 is side cross-section view of one embodiment of a
fluidic device 10. Fluidic device 10 includes a wall 22 which
encloses the sides and top of a reaction surface 12 to form a
reaction chamber 41. Reaction surface 12 includes an integral array
43. Coupled to the integral array may be an interface board 54.
Interface board 54 may interface the integral array 43 with
integrated circuits 50 via interconnect 55 and solder bumps 52.
[0096] In the embodiment of FIG. 8, interface board 54 may be
sandwiched between chamber 41 and integrated circuits 50. On one
side, interface board 54 may provide electrical signals (AC or DC)
to electrodes of integral array 43, while the other side of
interface board 54 may include pads for flip-chip mounting of
integrated circuits 50. Intermediate layers of interface board 54
may contain electrical leads, interconnects and vias, such as
interconnect 55 to transfer power and signals to and from
electrodes of integral array 43 and integrated circuits 50.
[0097] Interface board 54 may be fabricated using suitable PC-board
and flip chip technologies as is known in the art. Suitable
silk-screened or electroplated flip-chip solder bump techniques may
likewise be used. Alternatively, ink jet solder deposition may be
used as is known in the art.
[0098] FIG. 9 is a top view of an embodiment of a fluidic device
10. In the illustrated embodiment, fluidic device 10 is made up of
four distinct 8 by 8 integral arrays 43, forming a 16 by 16 array.
Under each 8 by 8 array may be situated an integrated circuit (not
shown in FIG. 9) that may provide control and signal processing to
electrodes of the integral array 43. The integral arrays may be
coupled to a circuit conducting pad 34 that may be coupled to an
interface conducting pad 36 by a bond wire 38 (shown only in one
quadrant). Connected to interface conducting pad 36 may be wire 42,
or another suitable connector such as a PC board connector, leading
to a computer or other suitable controlling device.
[0099] FIG. 9B is another top view of an embodiment of a fluidic
device 10. In this embodiment, many ports 15 are situated along
edges of fluidic device 10. These ports 15 may serve to inject
and/or collect packets 21 to/from reaction surface 12. Also
illustrated is a sensor 122 positioned adjacent a port 15. Such a
sensor is described in reference to FIG. 10 below.
[0100] FIG. 10 is a block diagram of a microfluidic processing
system 115. Processing system 115 may be designed to allow for
control of programmable dielectrophoretic array (PDA) 116 that
serves as the site for microfluidic interactions and may be
constructed in accordance with the present disclosure. In view of
its broad functionality, PDA 116 may serve a role, in the field of
fluidic processing, analogous to the role played by a Central
Processing Unit in the field of computers.
[0101] Coupled to PDA 116 are fluidic sensors 122. Fluidic sensors
122 may measure and monitor fluid products from, in, or on PDA 116.
For instance, fluidic sensors 122 may measure and identify reaction
products and may quantify reactions between packets. In one
embodiment, fluidic sensors 122 may include an optical microscope
or one or more sensors (chemical, electrochemical, electrical,
optical, or the like), but any other suitable monitoring device
known in the art may be substituted therewith. For example, fluidic
sensors 122 may be an electrochemical sensor that monitors the
presence and concentration of electroactive (redox-active)
molecules in a packet solution. An electrochemical sensor may take
the form of two or more microelectrodes. In a three-electrode
configuration, for example, electrodes may correspond to working,
reference, and counter electrodes. A packet to be analyzed may be
moved to be in contact with the three electrodes. A voltage signal
may be applied between the working and reference electrode, and the
current between the working and counter electrode may be monitored.
The voltage-current relationship allows for the determination of
the presence or absence, and concentration of electro-active
molecules in the packet solution. Also attached to PDA 116 may be
suitable material injection and extraction devices 120 coupled to
is appropriate inlet or outlet ports of PDA 116 (not shown in FIG.
10). Such devices may be any suitable structure allowing ingress to
and egress from PDA 116.
[0102] In electrical communication with PDA 116 may be PDA voltage
drivers 126 and dielectric position sensors 124. PDA voltage
drivers 126 may be adapted to drive electrodes within PDA 116 so
that an electric field may be established that sets up manipulation
forces that manipulate one or more packets of material within PDA
116 to promote microfluidic interactions. In one embodiment, PDA
voltage drivers 126 may include a signal generator and switching
network as described in relation to FIG. 7. Dielectric position
sensors 124 may measure positions of packets within PDA 116. In one
embodiment, dielectric position sensors 124 may include measuring
devices connected to appropriate sensors that may determine a
position of one or more packets of material by sensing, for
instance, a change in impedance between neighboring impedance
sensing electrodes within PDA 116 and by correlating that change in
impedance with a packet positioned adjacent the neighboring sensors
according to the teachings of the present disclosure.
[0103] Coupled to packet injection and extraction devices 120, PDA
voltage drivers 126, and dielectric position sensors 124 may be
computer interface 128. Computer interface 128 may be configured to
allow host computer 130 to interact with PDA 116. In one
embodiment, computer interface 128 may be a digital or analog card
or board that may analyze impedance data to obtain a packet
distribution map.
[0104] In the embodiment of FIG. 10, host computer 130 may be
coupled to computer interface 128 to provide for control of PDA
116. Host computer 130 may be coupled to position tracking agent
132 and to low-level control agent 134. Position tracking agent 132
may be adapted to store, process, and track positions of packets
within the fluidic processor PDA 116. Low-level control agent 134
may be configured to provide instructions to host computer 130 from
library interface 136 and software interface 138. Library interface
136 may hold various sets of subroutines for programmably
manipulating packets of materials on PDA 116. Software interface
138 that may allow for custom programming of instructions to be
executed by the fluidic processor PDA 116 to programmably
manipulate packets. Alternatively established programs of
manipulation instructions for specific fluid processing tests may
be read from stored data and executed by the PDA fluid processor
116.
[0105] FIG. 11 illustrates operation of the presently disclosed
method and apparatus. In FIG. 11, open squares represent electrodes
of an integral array. However, it is contemplated that the
description below applies equally well to a device utilizing
separate impedance sensing electrodes and driving electrodes.
[0106] In the illustrated embodiment, a packet 21a may be
introduced onto reaction surface 12 adjacent the location
represented by integral impedance sensor/electrode 201. The packet
may be compartmentalized in an immiscible partitioning medium (not
shown). The introduction of the packet may be accomplished using an
appropriate inlet port positioned adjacent to electrode 201.
Alternatively, a packet may be introduced adjacent electrode 201 by
applying an appropriate signal to electrode 201 to generate an
extraction force that may extract the packet from an inlet port or
from an injector directly onto reaction surface 12 and adjacent to
electrode 201.
[0107] Once positioned upon reaction surface 12, packet 21a may be
made to move along a predetermined path indicated by dashed line
250. A path may be specified in a number of different ways. In one
embodiment, a user may specifically define a path. For instance,
one may specify a path, through appropriate programming of a
controller or processing system, such as the one depicted numeral
250. Alternatively, a user may specify a starting position and an
ending position to define a path. For instance, a user may specify
that packet 21a is to be introduced adjacent electrode 201 and end
at a location adjacent electrode 215. Alternatively, one may
specify a starting and ending location with specific path
information in between. For instance, a user may specify a starting
position, an ending position, and a wavy path in between. As can be
seen from FIG. 11, the path may have any arbitrary shape and it may
be programmed in any number of ways.
[0108] To move packet 21a generally along the path, electrical
signals may be suitably switched to integral impedance
sensors/electrode pairs so that programmable manipulation forces
may be created that act upon packet 21a to propel it generally
along the specified path. As discussed earlier, the signals may be
varied in numerous ways to achieve the proper manipulation force.
In the illustrated embodiment, applying voltage signals to
electrode pairs 202 and 203 may create an attractive
dielectrophoretic force that moves packet 21a from electrode 201
towards electrode 203 generally along path 250. As packet 21a moves
generally along a specified path, the integral array may measure
impedances to map the position of the packet upon reaction surface
12. Knowing the position of a packet allows manipulation forces to
be directed at appropriate positions to achieve a desired
microfluidic processing task or interaction. In particular, knowing
a position of a packet allows an appropriate signal to be switched
to an appropriate electrode or electrode pair to generate a
manipulation force that further propels or interacts the packet
according to one or more instructions.
[0109] As packet 21a moves from electrode 201 towards electrode
203, the impedance between electrode 202 and electrode 203 may
change value, indicating that packet 21a is between, or partially
between, those two electrodes. The impedance may be measured as
described in FIG. 3. A controller or processing system (not shown
in FIG. 11) may register the location of packet 21a and may apply a
signal, for instance, to electrode pairs 204 and 205, creating an
attractive dielectrophoretic force which propels packet 21a towards
those electrodes generally along path 250. As the impedance between
electrode 204 and electrode 205 changes value, a controller or
processing system may apply a signal to electrodes 206 and 207 to
propel packet 21a along path 250. As packet 21a continues along
path 250, the impedance between electrode 206 and electrode 207 may
change value, indicating the presence of packet 21a adjacent that
location along the array. Thus, as packet 21a moves along path 250,
a controller or processing system may constantly monitor the
position of the packet by measuring impedance between electrode
pairs and adjust electrical signals to an appropriate electrode or
electrode pair (and hence, adjust manipulation forces) to continue
to propel the packet further along the specified path.
[0110] Measuring an impedance between pairs of electrodes not only
allows a position of a packet to be determined, but it also allows
for the determination of a location of an obstruction or blockage
upon reaction surface 12. For example, measuring the impedance
between electrodes 211 and 213 may indicate the presence of
obstruction 212. By noting the position of obstruction 212, a
controller or processing system may re-route one or more packets
around the obstruction so that no interference with microfluidic
processing interactions occurs. For example, if a path is specified
that passes through an area occupied by obstruction 212, a
controller or processing system may modify electrical signals to
propel a packet generally along the specified path while avoiding
the obstruction. For instance, a stronger or weaker signal may be
sent to one or more electrodes or electrode pairs near obstruction
212 to steer a packet clear of the blockage while still
maintaining, generally, the path that was originally specified, and
more particularly, the originally specified end point
[0111] A controller or processing system according to the presently
disclosed method and apparatus may be programmed to scan for
several obstructions and/or packets. Such a scan may build up a
distribution map, showing the location(s) of various packets and/or
obstructions on an entire reaction surface 12 or a portion thereof
Such a distribution map may be a virtual map, stored, for example,
in a computer memory or display. Turning again to FIG. 11,
impedances of all electrode pairs adjacent to path 250 may be
measured to determine if an obstruction blocks that path or if a
packet lies somewhere in that area. If the path is determined to be
clear (e.g., if all the electrode pairs show an impedance value
indicating a clear area), a packet may be safely propelled
generally along the path while avoiding any interactions with other
packets and/or obstructions. However, if an obstruction is
discovered, several different actions may be taken. In one
embodiment, the user may be notified that a blockage exists along
the specified path. The user may then specify a different path or
give another appropriate instruction. In another embodiment, the
controller or processing system may determine if the obstruction
may be avoided while still maintaining generally the same specified
path. If possible, electrical signals may be modified and delivered
to an electrode or electrode pairs to generate appropriate
electrical field distributions that set up proper manipulation
forces that will aid in avoiding the obstruction. Because, at least
in part, of this ability to constantly measure positions and
responses of packets during manipulation, a controller or
processing system may be capable of monitoring the integrity of
fluidic processing, reporting and correcting any errors that may
occur.
[0112] FIG. 11 also depicts how processing may be carried out on
two packets. In the illustrated embodiment, a second packet 21b
begins on reaction surface 12 near electrode 217. A second path,
path 260, may be specified that ends at electrode 219. As can be
seen, paths 250 and 260 may cross at interaction point 240. At
interaction point 240, the two packets may interact in many ways as
illustrated, for example, in FIG. 12. The interaction may include,
but is not limited to, fusing, merging, mixing, reacting, dividing,
splitting, or any combination thereof For instance, the two packets
may interact at interaction point 240 to form one or more
intermediate or final reaction products. Those products may be
manipulated in the same or in a similar manner as the two original
packets were manipulated.
[0113] FIG. 11 also depicts how maintenance may be performed upon
reaction surface 12. A maintenance packet 21c adapted to perform
maintenance upon reaction surface 12 may be introduced onto
reaction surface 12 by a maintenance port (not shown in FIG. 11). A
maintenance port may be similar to an inlet port in structure but
may be dedicated to the introduction of one or more maintenance
packets 21c designed specifically, for instance, to clean or
maintain reaction surface 12, a surface coating, or one or more
electrodes or sensors. Maintenance packet 21c may also react with
an obstruction in such a way as to remove that obstruction. As
illustrated, maintenance packet 21c may begin near electrode 241.
It may then be propelled along path 270, providing maintenance,
perhaps, to electrodes 242 and 243. Maintenance packet 21c may be
propelled back to a maintenance port, extracted from reaction
surface 12, and later used again, or it may discarded at an outlet
part.
[0114] FIG. 12 demonstrates several different possible fluidic
interactions that may be carried out using the presently disclosed
method and apparatus. In the illustrated embodiment, packets 21
(only one is labeled for convenience) reside upon a reaction
surface 12 having an integral array 43 (only one electrode is
labeled for convenience). In the top pane of FIG. 12, there is
shown an interaction in which a single packet is manipulated on the
reaction surface by moving the packet in a programmed fashion. In
the middle pane, two packets, starting at different locations upon
the reaction surface, are directed, via appropriate electrical
signals, to come together at a specified location (near the center
of the array) to fuse together, for example, to initiate a
reaction. The fused packet may be manipulated just as the original
packets were manipulated. For instance, the fused packet may be
moved to various locations or it may fuse again with another
packet(s). Shown in the bottom pane of FIG. 12 is a splitting
interaction. As shown, a single packet is subjected to different
programmable manipulation forces that cause the packet to split
into two distinct packets. Such an interaction may be accomplished
by, first, noting the position of the packet to be split, and then
by applying appropriate signals to electrode pairs to generate two
or more opposing forces that cause the packet to split apart.
[0115] FIG. 13 is a flowchart showing one embodiment of a method of
operation. A material may be introduced onto a reaction surface and
compartmentalized to form one or more packets in step 300. Multiple
materials may be introduced at different locations along reaction
surface 12 to form a plurality of packets. A path may be specified
as in step 310. The path may be designed to accomplish any type of
microfluidic processing, manipulation, or interaction. Different
reactions may be performed in serial or in parallel according to
different paths. Instructions governing such processing may be
embodied in the pseudo-code that may be routed through computer
interface 128 of FIG. 10. Illustrative code may read as
follows:
TABLE-US-00001 Example: AvidinActin.PSL Use inlet(1-3), outlet(1-2)
Inlet(1) is actin Inlet(2) is avidin Inlet(3) is enzyme Outlet(1)
is polymer Outlet(2) is waste Matrix(1,2) is accumulator Clean Do
Sactin = (Pull actin) // pull a new packet on the next Savidin =
(Pull avidin) // available matrix element next to Senzyme = (Pull
enzyme) // the inlets Move Sactin into accumulator // merges
components and enzyme Move Savidin into accumulator // in a single
packet Move Senzyme into accumulator Wait 1000ms ShiftRow
accumulator.row ,+1 // drag packet left into polymer outlet Move
0.5*accumulator into (2, accumulator.column)// drag half packet to
row 2 ShiftRow 2, + 1 // drag packet left into waste Loop Until
polymer.count = 10 // number of packet at polymer outlet = 10
Clean
[0116] In step 315, computer 80 of FIG. 6 or any other suitable
device may determine the next unit step along the path specified in
step 315. In other words, a path may be broken down into unit steps
and the next unit step or steps may be determined with respect to
the specified path. In step 320, a programmable manipulation force
is generated on reaction surface 12 through the use of any of the
mechanisms disclosed herein. The programmable manipulation force
may manipulate the one or more packets according to instructions
from a user. In step 330, the response(s) of the one or more
packets may be monitored. This step may include measuring an
impedance on the reaction surface as discussed herein. In
particular, one may determine whether the one or more packets moved
to where they were supposed to, or whether they interacted as
instructed. In step 340, it may be determined if the packet
movement was successful--that is, it may be determined whether the
packet ended up at a location corresponding to the unit step
determined in step 315.
[0117] If a packet movement was successful (i.e., if the packet
responded correctly to the programmable manipulation force(s)),
then it may be determined, by comparison with the specified path,
whether or not the packet destination has been reached. In other
words, it may be determined if the packet has moved to the end
location of the specified path. If the destination has not been
reached, the next unit step movement may be determined at step 315
and steps 320, 330, 340, and 365 may be repeated. If the
destination has been reached, it may be determined whether another
packet is to be manipulated in step 370. This step may include a
user prompt. If no further packets are to be manipulated, it may be
determined whether fluidic processing is complete in step 380. If
yes, the process may be ended at step 390. Step 390 may include the
collecting of one or more packets, further analysis, throwing away
of the reaction surface, or any procedure described herein. If the
processing is not complete, the next step of processing may be
determined in step 395. The next step may entail, for example, the
introduction of another packet, the specification of another path,
or any other step of FIG. 13.
[0118] If a packet manipulation is unsuccessful (i.e., if the
applied programmable manipulation force(s) did not produce a
desired interaction or movement along a specified path as indicated
by step 340), one may locate an obstruction upon the reaction
surface as indicated in step 350 and as taught herein. After
locating any obstructions, a new, modified path may be determined
or specified as indicated by step 360, leading to step 310.
[0119] As mentioned with relation to FIG. 1, the present disclosure
contemplates that many different types of forces may be utilized as
a manipulation force for promoting fluidic interactions among
packets of material on a reaction surface. Specifically, suitable
forces other than dielectrophoresis include electrophoretic forces,
optical forces, mechanical forces, or any combination thereof.
Below are discussed embodiments of the present disclosure dealing
with electrophoretic and optical manipulation forces.
Programmable Electrophoretic Array (PEA)
[0120] A fluidic processing system incorporating a programmable
electrophoretic array may be constructed according to the present
disclosure. As used herein, "programmable electrophoretic array"
(PEA) refers to an electrode array whose individual elements can is
be addressed with DC, pulsed, or low frequency AC electrical
signals (typically, less than about 10 kHz) electrical signals. The
addressing of electrode elements with electrical signals initiates
different field distributions and generates electrophoretic
manipulation forces that trap, repel, transport or perform other
manipulations upon charged packets on and above the electrode
plane. By programmably addressing electrode elements within the
array with electrical signals, electric field distributions and
electrophoretic manipulation forces acting upon charged packets may
be programmable so that packets may be manipulated along
arbitrarily chosen or predetermined paths. A PEA may utilize
electrophoretic forces in DC or low-frequency (typically, less than
about 10 kHz) AC electrical fields. Such electrophoretic forces may
be used instead of, or in addition to, another manipulation forces
such as dielectrophoresis.
[0121] Negative or positive charges may be induced or injected into
fluid packets. The charged packets may be moved or manipulated by
electrophoretic forces generated by an electrode array fabricated
on an inner surfaces of a chamber in accordance with this
disclosure. The electrode array, termed a programmable
electrophoretic array (PEA), may consist of uniformly or
non-uniformly spaced electrode elements. Individual electrode
elements may be independently addressable with DC, pulsed, or low
frequency AC electrical signals (<about 10 kHz). Characteristic
dimensions of individual electrode elements may be of any size but,
in one embodiment, may lie between 0.2 micron and 10 mm. Individual
electrode elements may take similar or different geometrical forms
such as squares, circles, diamonds, or other shapes. Programmably
switchable electrical signals may be applied to individual
electrode elements so that a programmable electrical field
distribution may be generated. Such a distribution may impose
electrophoretic forces to trap, repel, transport or manipulate
charged packets in a partitioning medium. Further, electrical
signals may be applied to such an array so that a packet may be
broken down to two or more packets. The programmability of a PEA
may be reflected in the fact that the electric field distributions
and electrophoretic forces acting on charged packets may be
programmable so that charged packets may be trapped or repelled or
transported along arbitrarily chosen paths in the partitioning
medium, and that a PEA may be programmed to perform different
reactions in series or in parallel where different manipulation
protocols of packets (differing in size, number, and/or reagent
type concentration) may be required. As with PDA surface
modification, if a dielectric layer coating is applied to the
surface of a PEA to modify interaction forces between packets
reaction surfaces, the dielectric layer may be made sufficiently
thin (typically 2 nm to 1 micron) to allow for electric field
penetration.
Optical Manipulation
[0122] Optical tweezers (which may consist of a focused laser beam
with a light intensity gradient) may be also be used for trapping
and manipulating packets of material. Optical manipulation requires
that the refractive indices of the packets be different from that
of their suspending medium, for instance, a partitioning medium as
described herein. As light passes through one or more packets, it
may induce fluctuating dipoles. Those dipoles may interact with
electromagnetic field gradients, resulting in optical forces
directed towards or away from the brighter region of the light. If
their refractive indices are higher than that of the partitioning
medium, packets may be trapped in a bright region, and when the
laser light moves with respect to the partitioning medium, packets
may follow the light beam, allowing for optical manipulation
forces. Conversely, if the packets have refractive indices smaller
than their partitioning medium, they will experience forces
directing them away from bright regions.
[0123] Therefore, if packets have different refractive indexes from
that of the partitioning medium (e.g., water packets in air or
oil), optical tweezers may exert forces on them. Therefore, to
manipulate and interact packets, a microscope or other optical
system incorporating one or more laser tweezers may be used. A
chamber containing a partitioning medium in accordance with the
present disclosure may be placed into such an optical system.
Following the introduction of packets of material into the chamber,
laser tweezers may be used to trap packets. By moving the focal
point of the optical tweezers with respect to the partitioning
medium (e.g., moving a stage holding the thin chamber containing
the partitioning medium whilst fixing the position of laser
tweezers and/or by focusing the laser beam to different depths in
the partitioning medium), packets may be manipulated as described
herein. Through the use of apparatus such as a
computer-controllable, multi-axis translation stage, the movement
of the optical tweezers with respect to the suspending medium may
be programmed or automatically controlled. Thus the optical tweezer
may be moved, with respect to the medium, along any arbitrarily
chosen or predetermined paths. By doing so, packets under the
influences of the optical tweezers may be manipulated along any
arbitrarily chosen or predetermined paths.
EXAMPLE 1
[0124] Aqueous materials have been compartmentalized to form
packets using hydrophobic liquids as a partitioning medium.
Partitioning mediums so used have included decane, bromodocane,
mineral oil, and 3 in 1.TM. oil. Packets have been formed by
briefly sonicating about 3 milliliters of the hydrophobic liquid to
which had been added 20 to 50 microliters of aqueous medium.
Aqueous media tested have included deionized water, tap water
(electrical conductivity of about 40 mS/m) and phosphate buffered
saline (PBS) solution.
EXAMPLE 2
[0125] Aqueous packets suspended in mineral oil, bromodoecane and 3
in 1.TM. oil have been collected by dielectrophoresis by applying
sinusoidal signals to gold-on-glass electrode arrays having 20, 80
and 160 micron spacing, respectively. The 20-micron electrode array
consisted of parallel line electrodes (20 microns in width and
spacing). The 80 and 160 micron electrode arrays were of the
interdigitated, castellated geometries. Aqueous packets were
collected at electrode edges or tips when AC voltage signals
between 100 Hz and 20 MHz were applied. Applied voltages were from
10 to 100 V peak-to-peak. The formation of pearl-chains of water
packets has also been observed.
EXAMPLE 3
[0126] Aqueous packets in hydrophobic suspension have been brought
together and fused under the influence of dielectrophoretic forces
on the same electrode arrays used in Example 2.
EXAMPLE 4
[0127] Packets have been moved from one electrode element to
another under influence of dielectrophoretic forces when the AC
electrical field is switched on an addressable array of parallel
line electrodes having 20 micron width and spacing.
EXAMPLE 5
[0128] Sensitive AC impedance monitors have been built for use with
microelectrode arrays. Such monitors may provide for sensitive
dielectric sensing of packet positions.
[0129] While the present disclosure may be adaptable to various
modifications and alternative forms, specific embodiments have been
shown by way of example and described herein. However, it should be
understood that the present disclosure is not intended to be
limited to the particular forms disclosed. Rather, it is to cover
all modifications, equivalents, and alternatives falling within the
spirit and scope of the disclosure as defined by the appended
claims. Moreover, the different aspects of the disclosed apparatus
and methods may be utilized in various combinations and/or
independently. Thus the invention is not limited to only those
combinations shown herein, but rather may include other
combinations.
* * * * *