Comparison of Specifications in U.S. Patent No. 6,506,609 (Ex. 1006) and U.S.
`Application No. 09/569,747 (Ex. 1018)
`U.S. Application No. 09/569,747 (Ex. 1018)
`U.S. Patent No. 6,506,609 (Ex. 1006)
`A variety of cell-based assays are of
`A variety of cell-based assays are of considerable
`considerable commercial relevance in screening
`commercial relevance in screening for
`for modulators of cell-based activity. For
`modulators of cell-based activity. For example,
`example, compounds which affect cell death
`compounds which affect cell death can have
`can have profound biological activities and are
`profound biological activities and are desirably
`desirably screened for in cell-based assays. Cell
`screened for in cell-based assays. Cell death has
`death has become recognized as a physiological
`become recognized as a physiological process
`process important in normal development,
`important in normal development, hormonal
`hormonal regulation of various tissues, and,
`regulation of various tissues, and, e.g., in
`e.g., in regulation of the receptor repertoires of
`regulation of the receptor repertoires of both T
`both T and B lymphocytes. The finding that a
`and B lymphocytes. The finding that a pattern of
`pattern of morphological changes is common to
`morphological changes is common to many
`many examples of programmed cell death (or
`examples of programmed cell death (or PCD) led
`PCD) led to the suggestion of a common
`to the suggestion of a common mechanism, and
`mechanism, and the term "apoptosis" was
`the term “apoptosis” was defined to include both
`defined to include both the morphological
`the morphological features and the mechanism
`features and the mechanism common to such
`common to such programmed cell death (Kerr et
`programmed cell death (Kerr et al., Br. J.
`al., Br. J. Cancer 26:239). This concept was
`Cancer 26:239). This concept was extended by
`extended by the finding that nuclear DNA
`the finding that nuclear DNA fragmentation
`fragmentation correlates well with apoptotic
`correlates well with apoptotic morphology
`morphology (Arends et al. Am. J.
`(Arends et al., Am. J. Pathol. 136:593 (1990)),
`Pathol. 136:593 (1990)), and the scientific
`and the scientific literature contains many
`literature contains many examples of PCD
`examples of PCD accompanied by these
`accompanied by these features. There are also
`features. There are also clear examples of PCD
`clear examples of PCD in the absence of
`in the absence of apoptotic morphology or
`apoptotic morphology or DNA fragmentation
`DNA fragmentation (Clarke, Anat. Embryl.
`(Clarke, Anat. Embryl. 181:195 (1990), Martin et
`181:195 (1990), Martin et al, J. Cell Biol.
`al, J. Cell Biol. 106:829 (1988), and Ishigami et
`106:829 (1988), and Ishigami et al., J. Immunol
`al., J. Immunol. 148:360 (1992)).
`148:360 (1992)).
`Cell-based assay systems model relevant
`biological phenomena, and have generally been
`widely adopted as screening assays, e.g., when
`screening for a compound's effect(s) on
`apoptosis or other biological phenomena.
`Pioneering technology providing cell- and other
`particle-based microscale assays are set forth in
`Parce et al. "High Throughput Screening Assay
`Systems in Microscale Fluidic Devices" WO
`98/00231; in PCT/US00/04522, filed February
`22, 2000, entitled "Manipulation of
`Microparticles In Microfluidic Systems," by
`
`Cell-based assay systems model relevant
`biological phenomena, and have generally been
`widely adopted as screening assays, e.g., when
`screening for a compound's effect(s) on apoptosis
`or other biological phenomena. Pioneering
`technology providing cell- and other particle-
`based microscale assays are set forth in Parce al.
`“High Throughput Screening Assay Systems in
`Microscale Fluidic Devices” WO 98/00231; in
`PCT/US00/04522, filed FebruaryFeb. 22, 2000,
`entitled “Manipulation of Microparticles In
`Microfluidic Systems,” by Mehta et al.; and in
`
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`Mehta et al.; and in PCT/US00/04486, filed
`February 22, 2000, entitled "Devices and
`Systems for Sequencing by Synthesis," by
`Mehta et al.
`Other cell-based assays include various
`methods for the preparative or analytic sorting
`of different types of cells. For example, cell
`panning generally involves attaching an
`appropriate antibody or other cell-specific
`reagent to a solid support and then exposing the
`solid support to a heterogeneous cell sample.
`Cells possessing, e.g., the corresponding
`membrane-bound antigen will bind to the
`support, leaving those lacking the appropriate
`antigenic determinant to be washed away. Other
`well-known sorting methods include those
`using fluorescence-activated cell sorters
`("FACSs"). FACSs for use in sorting cells and
`certain subcellular components such as
`molecules of DNA have been proposed in, e.g.,
`Fu, A.Y. et al. (1999) "A Microfabricated
`Fluorescence-Activated Cell Sorter," Nat.
`Biotechnol. 17:1109-1111; Unger, M., et al.
`(1999) "Single Molecule Fluorescence
`Observed with Mercury Lamp Illumination,"
`Biotechniques 27:1008-1013; and Chou, H.P. et
`al. (1999) "A Microfabricated Device for Sizing
`and Sorting DNA Molecules," Proc. Nat'l.
`Acad. Sci. 96:11-13. These sorting techniques
`utilizing generally involve focusing cells or
`other particles by flow channel geometry.
`While cell-based assays are generally preferred
`in certain microscale screening applications,
`certain of these assays are difficult to adapt to
`conventional notions of high-throughput or
`ultra-high-throughput screening assay systems.
`For example, one difficulty in flowing assay
`systems is that, during pressure-based flow of
`fluids in channels, non-uniform flow velocities
`are experienced. Faster fluid and material flow
`is observed in the center of a moving fluid
`stream than on the edge of a moving fluid
`stream. This non-uniform flow velocity reduces
`throughput for flowing assays, because assay
`runs have to be spaced well apart in the fluid
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`PCT/USPCTUS00/04486, filed FebruaryFeb.
`22, 2000, entitled “Devices and Systems for
`Sequencing by Synthesis,” by Mehta et al.
`
`Other cell-based assays include various methods
`for the preparative or analytic sorting of different
`types of cells. For example, cell panning
`generally involves attaching an appropriate
`antibody or other cell-specific reagent to a solid
`support and then exposing the solid support to a
`heterogeneous cell sample. Cells possessing, e.g.,
`the corresponding membrane-bound antigen will
`bind to the support, leaving those lacking the
`appropriate antigenic determinant to be washed
`away. Other well-known sorting methods include
`those using fluorescence-activated cell sorters
`(“FACSs”). FACSs for use in sorting cells and
`certain subcellular components such as
`molecules of DNA have been proposed in, e.g.,
`Fu, A. Y. et al. (1999) “A Microfabricated
`Fluorescence-Activated Cell Sorter,” Nat.
`Biotechnol. 17:1109-1111; Unger, M., et al.
`(1999) “Single Molecule Fluorescence Observed
`with Mercury Lamp Illumination,”
`Biotechniques 27:1008-1013; and Chou, H. P. et
`al. (1999) “A Microfabricated Device for Sizing
`and Sorting DNA Molecules,” Proc. Nat'l. Acad.
`Sci. 96:11-13. These sorting techniques utilizing
`generally involve focusing cells or other particles
`by flow channel geometry.
`
`While cell-based assays are generally preferred
`in certain microscale screening applications,
`certain of these assays are difficult to adapt to
`conventional notions of high-throughput or ultra-
`high-throughput screening assay systems. For
`example, one difficulty in flowing assay systems
`is that, during pressure-based flow of fluids in
`channels, non-uniform flow velocities are
`experienced. Faster fluid and material flow is
`observed in the center of a moving fluid stream
`than on the edge of a moving fluid stream. This
`non-uniform flow velocity reduces throughput
`for flowing assays, because assay runs have to be
`spaced well apart in the fluid stream to prevent
`
`ABS Global, Inc. and Genus plc – Ex. 1007, p. 2
`
`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`stream to prevent overlap of materials moving
`at different velocities.
`Accordingly, it would be advantageous to
`provide mechanisms for facilitating cell-based
`assays, including cell sorting techniques,
`especially in microscale systems. Additional
`microscale assays directed at subcellular
`components, such as nucleic acids would also
`be desirable. The present invention provides
`these and other features which will become
`clear upon consideration of the following.
`The present invention relates to methods of
`focusing particles in microchannels, e.g., to
`improve assay throughput, to sort particles, to
`count particles, or the like. In the methods of
`the invention, cells and other particles are
`focused in the center of, to one side of, or in
`other selected regions of microscale channels,
`thereby avoiding, e.g., the above noted
`difficulties inherent in pressure-based flow of
`particles. Furthermore, the device structures of
`the present invention are optionally integrated
`with other microfluidic systems. Other reactions
`or manipulations involving cells, other
`particles, or fluids upstream of the detection
`zone are also optionally performed, e.g.,
`monitoring drug interactions with cells or other
`particles.
`In one aspect, the invention provides methods
`of providing substantially uniform flow velocity
`to particles flowing in a first microchannel. In
`the methods, the particles are optionally flowed
`in the microchannel, e.g., using pressure- based
`flow, in which the particles flow with a
`substantially non-uniform flow velocity. Prior
`to performing the flowing step, the particles are
`optionally sampled with at least one capillary
`element, e.g., by dipping the capillary element
`into a well containing the particles on a
`microcell plate and drawing the particles into,
`e.g., reservoirs, microchannels, or other
`chambers of the device. The particles (e.g., a
`cell, a set of cells, a microbead, a set of
`microbeads, a functionalized microbead, a set
`of functionalized microbeads, a molecule, a set
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`overlap of materials moving at different
`velocities.
`Accordingly, it would be advantageous to
`provide mechanisms for facilitating cell-based
`assays, including cell sorting techniques,
`especially in microscale systems. Additional
`microscale assays directed at subcellular
`components, such as nucleic acids would also be
`desirable. The present invention provides these
`and other features which will become clear upon
`consideration of the following.
`The present invention relates to methods of
`focusing particles in microchannels, e.g., to
`improve assay throughput, to sort particles, to
`count particles, or the like. In the methods of the
`invention, cells and other particles are focused in
`the center of, to one side of, or in other selected
`regions of microscale channels, thereby avoiding,
`e.g., the above noted difficulties inherent in
`pressure-based flow of particles. Furthermore,
`the device structures of the present invention are
`optionally integrated with other microfluidic
`systems. Other reactions or manipulations
`involving cells, other particles, or fluids
`upstream of the detection zone are also
`optionally performed, e.g., monitoring drug
`interactions with cells or other particles.
`
`In one aspect, the invention provides methods of
`providing substantially uniform flow velocity to
`particles flowing in a first microchannel. In the
`methods, the particles are optionally flowed in
`the microchannel, e.g., using pressure-based
`flow, in which the particles flow with a
`substantially non-uniform flow velocity. Prior to
`performing the flowing step, the particles are
`optionally sampled with at least one capillary
`element, e.g., by dipping the capillary element
`into a well containing the particles on a microcell
`plate and drawing the particles into, e.g.,
`reservoirs, microchannels, or other chambers of
`the device. The particles (e.g., a cell, a set of
`cells, a microbead, a set of microbeads, a
`functionalized microbead, a set of functionalized
`microbeads, a molecule, a set of molecules, etc.)
`
`ABS Global, Inc. and Genus plc – Ex. 1007, p. 3
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`of molecules, etc.) are optionally focused
`horizontally and/or vertically in the first
`microchannel to provide substantially uniform
`flow velocity to the particles in the first
`microchannel. Particles are optionally focused
`using one or more fluid direction components,
`e.g. a fluid pressure force capillary force
`modulator, an electrokinetic force modulator, a
`capillary force modulator, a fluid wicking
`element, or the like). Additional options include
`sorting, detecting or otherwise manipulating the
`focused particles.
`The particles are horizontally focused in the
`microchannel, e.g., by introducing a low density
`fluid and a high density fluid into the
`microchannel, causing the particles to be
`focused in an intermediate density fluid present
`between the high density fluid and the low
`density fluid. The particles are also optionally
`focused in a top or a bottom portion of the
`microchannel by introducing a high or a low
`density fluid into the microchannel with the
`flowing particles. The particles are vertically or
`horizontally focused in the microchannel, e.g.,
`by simultaneously introducing fluid flow from
`two opposing microchannels into the first
`microchannel during flow of the particles in the
`first channel. Vertical focusing is also
`optionally achieved to one side of a
`microchannel by simultaneously introducing
`fluid flow from, e.g., a second microchannel
`into the first microchannel during flow of the
`particles in the first microchannel.
`In another aspect, the invention also provides
`particle washing or exchange techniques. For
`example, focused cells or other particles are
`optionally washed free of diffusible material by
`introducing a diluent into the first microchannel
`from at least a second channel and removing the
`resulting diluted diffused product comprising
`diluent mixed with the diffusible material
`through at least a third microchannel.
`Alternating arrangements of diluent input and
`diffused product output channels are also
`optionally used to further wash the particles.
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`are optionally focused horizontally and/or
`vertically in the first microchannel to provide
`substantially uniform flow velocity to the
`particles in the first microchannel. Particles are
`optionally focused using one or more fluid
`direction components, e.g. a fluid pressure force
`capillary force modulator, an electrokinetic force
`modulator, a capillary force modulator, a fluid
`wicking element, or the like). Additional options
`include sorting, detecting or otherwise
`manipulating the focused particles.
`
`The particles are horizontally focused in the
`microchannel, e.g., by introducing a low density
`fluid and a high density fluid into the
`microchannel, causing the particles to be focused
`in an intermediate density fluid present between
`the high density fluid and the low density fluid.
`The particles are also optionally focused in a top
`or a bottom portion of the microchannel by
`introducing a high or a low density fluid into the
`microchannel with the flowing particles. The
`particles are vertically or horizontally focused in
`the microchannel, e.g., by simultaneously
`introducing fluid flow from two opposing
`microchannels into the first microchannel during
`flow of the particles in the first channel. Vertical
`focusing is also optionally achieved to one side
`of a microchannel by simultaneously introducing
`fluid flow from, e.g., a second microchannel into
`the first microchannel during flow of the
`particles in the first microchannel.
`
`In another aspect, the invention also provides
`particle washing or exchange techniques. For
`example, focused cells or other particles are
`optionally washed free of diffusible material by
`introducing a diluent into the first microchannel
`from at least a second channel and removing the
`resulting diluted diffused product comprising
`diluent mixed with the diffusible material
`through at least a third microchannel.
`Alternating arrangements of diluent input and
`diffused product output channels are also
`optionally used to further wash the particles. For
`
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`For example, in one aspect the methods of the
`invention include simultaneously introducing
`the diluent into the first microchannel from the
`second microchannel and a fourth
`microchannel, where the second and fourth
`microchannel intersect the first microchannel at
`a common intersection region. Optionally, the
`methods include sequentially introducing the
`diluent into the first microchannel from the
`second microchannel and a fourth
`microchannel, wherein the second and fourth
`microchannels intersect the first microchannel
`at an offset intersection region. The diffused
`product is typically removed through the third
`microchannel and a fifth microchannel, which
`third and fifth microchannels intersect the first
`microchannel at a common intersection region.
`In further washing steps, the diluent is
`introduced through sixth and seventh
`microchannels which intersect the first
`microchannel at a common intersection. The
`resulting further diluted diffused product is
`removed through eighth and ninth
`microchannels, which intersect the first
`microchannel at a common intersection. Diluent
`is optionally introduced into the first
`microchannel by pressure or electrokinetic
`flow.
`In one preferred assay of the invention, the
`particles are cells and the method includes
`performing a TUNEL assay or an Annexin-V
`assay on the cells in the channel to measure
`apoptosis.
`Integrated systems for performing the above
`methods, including the particle sorting
`embodiments, are also provided.
`An integrated system for providing
`substantially uniform flow velocity to flowing
`members of at least one particle population in a
`microfluidic device optionally includes a body
`structure that includes at least a first
`microchannel disposed therein. A first fluid
`direction component (e.g., a fluid pressure force
`modulator) is typically coupled to the first
`microchannel for inducing flow of a fluidic
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`example, in one aspect the methods of the
`invention include simultaneously introducing the
`diluent into the first microchannel from the
`second microchannel and a fourth microchannel,
`where the second and fourth microchannel
`intersect the first microchannel at a common
`intersection region. Optionally, the methods
`include sequentially introducing the diluent into
`the first microchannel from the second
`microchannel and a fourth microchannel,
`wherein the second and fourth microchannels
`intersect the first microchannel at an offset
`intersection region. The diffused product is
`typically removed through the third
`microchannel and a fifth microchannel, which
`third and fifth microchannels intersect the first
`microchannel at a common intersection region.
`In further washing steps, the diluent is introduced
`through sixth and seventh microchannels which
`intersect the first microchannel at a common
`intersection. The resulting further diluted
`diffused product is removed through eighth and
`ninth microchannels, which intersect the first
`microchannel at a common intersection. Diluent
`is optionally introduced into the first
`microchannel by pressure or electrokinetic flow.
`
`In one preferred assay of the invention, the
`particles are cells and the method includes
`performing a TUNEL assay or an Annexin-V
`assay on the cells in the channel to measure
`apoptosis.
`Integrated systems for performing the above
`methods, including the particle sorting
`embodiments, are also provided.
`An integrated system for providing substantially
`uniform flow velocity to flowing members of at
`least one particle population in a microfluidic
`device optionally includes a body structure that
`includes at least a first microchannel disposed
`therein. A first fluid direction component (e.g., a
`fluid pressure force modulator) is typically
`coupled to the first microchannel for inducing
`flow of a fluidic material that includes the
`
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`material that includes the members of the at
`least one particle population in the first
`microchannel. The first fluid direction
`component generally induces non-uniform
`flow. A source of at least one fluidic material is
`optionally fluidly coupled to the first
`microchannel. The system also optionally
`includes at least a second microchannel that
`intersects the first microchannel for introducing
`at least one fluid into the first microchannel to
`horizontally or vertically focus the members of
`the at least one particle population in the first
`microchannel. The at least one fluid is
`optionally introduced using a second fluid
`direction component that includes one or more
`of a fluid pressure force modulator, an
`electrokinetic force modulator, a capillary force
`modulator, a fluid wicking element, or the like.
`At least one flow control regulator for
`regulating flow of the fluidic material or the
`fluid in the first or second microchannel is also
`optionally provided. A computer including an
`instruction set directing simultaneous flow of
`the members of the at least one particle
`population in the first microchannel and
`simultaneous introduction of the at least one
`fluid from the second microchannel into the
`first microchannel is optionally also operably
`coupled to a fluid movement system for
`directing flow of materials in the
`microchannels.
`As a further option, this integrated system
`additionally includes at least a third
`microchannel which intersects the first
`microchannel in an intersection region common
`to the second microchannel. The flow control
`regulator of this system optionally further
`regulates flow of the at least one fluid in the
`second and the third microchannels. In this
`embodiment, the computer typically also
`includes an instruction set for simultaneously
`flowing fluids from the second and third
`microchannels into the first microchannel.
`In particle washing systems, typically, at least
`fourth and fifth channels which intersect the
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`members of the at least one particle population in
`the first microchannel. The first fluid direction
`component generally induces non-uniform flow.
`A source of at least one fluidic material is
`optionally fluidly coupled to the first
`microchannel. The system also optionally
`includes at least a second microchannel that
`intersects the first microchannel for introducing
`at least one fluid into the first microchannel to
`horizontally or vertically focus the members of
`the at least one particle population in the first
`microchannel. The at least one fluid is optionally
`introduced using a second fluid direction
`component that includes one or more of a fluid
`pressure force modulator, an electrokinetic force
`modulator, a capillary force modulator, a fluid
`wicking element, or the like. At least one flow
`control regulator for regulating flow of the
`fluidic material or the fluid in the first or second
`microchannel is also optionally provided. A
`computer including an instruction set directing
`simultaneous flow of the members of the at least
`one particle population in the first microchannel
`and simultaneous introduction of the at least one
`fluid from the second microchannel into the first
`microchannel is optionally also operably coupled
`to a fluid movement system for directing flow of
`materials in the microchannels.
`
`As a further option, this integrated system
`additionally includes at least a third
`microchannel which intersects the first
`microchannel in an intersection region common
`to the second microchannel. The flow control
`regulator of this system optionally further
`regulates flow of the at least one fluid in the
`second and the third microchannels. In this
`embodiment, the computer typically also
`includes an instruction set for simultaneously
`flowing fluids from the second and third
`microchannels into the first microchannel.
`In particle washing systems, typically, at least
`fourth and fifth channels which intersect the first
`
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`first microchannel at a common intersection
`downstream of the second and third
`microchannels are provided. The computer
`further includes an instruction set for
`simultaneously flowing material from the first
`microchannel into the fourth and fifth
`microchannels. Sixth and seventh
`microchannels which intersect the first
`microchannel at a common intersection
`downstream of the fourth and fifth
`microchannels, with the computer further
`comprising an instruction set for simultaneously
`flowing material from the sixth and seventh
`microchannels into the first microchannel are
`optionally provided. Similarly, eighth and ninth
`microchannels which intersect the first
`microchannel at a common intersection
`downstream of the sixth and seventh
`microchannels, the computer further including
`an instruction set for simultaneously flowing
`material from the first microchannel into the
`eighth and ninth microchannels are optionally
`provided.
`The integrated system optionally includes
`sources for any reagent or particle used in the
`methods noted above, such as one or more
`sources of terminal deoxynucleotide
`transferase, one or more sources of one or more
`fluorescein labeled nucleotides or other labeled
`polynucleotides, one or more sources of
`Annexin V, one or more sources of an
`AnnexinV-biotin conjugate, one or more
`sources of a DNA dye, one or more sources of
`Campthotecin, one or more sources of Calcein-
`AM, one or more sources of a control cell, one
`or more sources of a test cell, etc.
`Signal detector(s), mounted proximal to the first
`microchannel for detecting a detectable signal
`produced by one or more of the members of the
`at least one particle population in the
`microchannel are typically provided in the
`integrated systems of the invention. The
`detector also optionally includes, e.g., a
`fluorescent excitation source and a
`fluorescent emission detection element.
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`microchannel at a common intersection
`downstream of the second and third
`microchannels are provided. The computer
`further includes an instruction set for
`simultaneously flowing material from the first
`microchannel into the fourth and fifth
`microchannels. Sixth and seventh microchannels
`which intersect the first microchannel at a
`common intersection downstream of the fourth
`and fifth microchannels, with the computer
`further comprising an instruction set for
`simultaneously flowing material from the sixth
`and seventh microchannels into the first
`microchannel are optionally provided. Similarly,
`eighth and ninth microchannels which intersect
`the first microchannel at a common intersection
`downstream of the sixth and seventh
`microchannels, the computer further including an
`instruction set for simultaneously flowing
`material from the first microchannel into the
`eighth and ninth microchannels are optionally
`provided.
`
`The integrated system optionally includes
`sources for any reagent or particle used in the
`methods noted above, such as one or more
`sources of terminal deoxynucleotide transferase,
`one or more sources of one or more fluorescein
`labeled nucleotides or other labeled
`polynucleotides, one or more sources of Annexin
`V, one or more sources of an AnnexinV-biotin
`conjugate, one or more sources of a DNA dye,
`one or more sources of Campthotecin, one or
`more sources of Calcein-AM, one or more
`sources of a control cell, one or more sources of
`a test cell, etc.
`Signal detector(s), mounted proximal to the first
`microchannel for detecting a detectable signal
`produced by one or more of the members of the
`at least one particle population in the
`microchannel are typically provided in the
`integrated systems of the invention. The detector
`also optionally includes, e.g., a fluorescent
`excitation source and a fluorescent emission
`detection element. Optionally, the computer is
`
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`

`

`U.S. Application No. 09/569,747 (Ex. 1018)
`Optionally, the computer is operably linked
`to the signal detector and has an instruction
`set for converting detected signal information
`into digital data.
`The integrated system of the present invention
`is also optionally used to sort the members of a
`particle population (e.g., a cell, a set of cells, a
`microbead, a set of microbeads, a
`functionalized microbead, a set of
`functionalized microbeads, a molecule, a set of
`molecules, or the like). In this embodiment, the
`integrated system typically additionally
`includes a third and a fourth microchannel
`which intersect the first microchannel
`downstream from the intersection of the second
`microchannel with the first microchannel. The
`fourth microchannel also generally intersects
`the first microchannel downstream from the
`intersection of the third microchannel with the
`first microchannel. The flow control regulator
`of this system optionally further regulates flow
`of the at least one fluid in the third or the fourth
`microchannels. Furthermore, the signal detector
`typically detects a detectable signal produced
`by a selected member of the particle population
`between the intersections of the second and the
`third microchannels with the first microchannel.
`In this particle sorting embodiment, the
`computer is optionally operably linked to the
`first or other fluid direction component(s), the
`flow control regulator, and the signal detector.
`Additionally, the instruction set typically directs
`simultaneous introduction of the at least one
`fluid from the third microchannel into the first
`microchannel to horizontally or vertically focus
`the selected member of the particle population
`such that the selected member is directed into
`the fourth microchannel in response to the
`detectable signal produced by the selected
`member. Optionally, the instruction set further
`directs simultaneous introduction of the at least
`one fluid from the third microchannel by
`activating a heating element (e.g., a Joule
`heating electrode, a conductively coated
`microchannel portion, etc.) disposed within the
`
`U.S. Patent No. 6,506,609 (Ex. 1006)
`operably linked to the signal detector and has an
`instruction set for converting detected signal
`information into digital data.
`
`The integrated system of the present invention is
`also optionally used to sort the members of a
`particle population (e.g., a cell, a set of cells, a
`microbead, a set of microbeads, a functionalized
`microbead, a set of functionalized microbeads, a
`molecule, a set of molecules, or the like). In this
`embodiment, the integrated system typically
`additionally includes a third and a fourth
`microchannel which intersect the first
`microchannel downstream from the intersection
`of the second microchannel with the first
`microchannel. The fourth microchannel also
`generally intersects the first microchannel
`downstream from the intersection of the third
`microchannel with the first microchannel. The
`flow control regulator of this system optionally
`further regulates flow of the at least one fluid in
`the third or the fourth microchannels.
`Furthermore, the signal detector typically detects
`a detectable signal produced by a selected
`member of the particle population between the
`intersections of the second and the third
`microchannels with the first microchannel.
`In this particle sorting embodiment, the computer
`is optionally operably linked to the first or other
`fluid directions component(s), the flow control
`regulator, and the signal detector. Additionally,
`the instruction set typically directs simultaneous
`introduction of the at least one fluid from the
`third microchannel into the first microchannel to
`horizontally or vertically focus the selected
`member of the particle population such that the
`selected member is directed into the fourth
`microchannel in response to the detectable signal
`produced by the selected member. Optionally,
`the instruction set further directs simultaneous
`introduction of the at least one fluid from the
`th