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Claims  |
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We claim:
1. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and in fluid
communication and crossing said analysis channel at a first intersection;
a plurality of separate sample sources in fluid communication with said
sample loading channel, whereby there is at least one of said plurality of
separate sample sources in fluid communication with said sample loading
channel on each side of said first intersection; and
first and second load/waste channels disposed in said substrate, each of
said first and second load/waste channels intersecting said sample loading
channel at second and third intersections, respectively, said second and
third intersections being on different sides of said first intersection.
2. The microfluidic system of claim 1, wherein said second and third
intersections are each within 5 mm of said first intersection.
3. The microfluidic system of claim 1, wherein said second and third
intersections are each within 2 mm of said first intersection.
4. The microfluidic system of claim 1, wherein said second and third
intersections are each within 1 mm of said first intersection.
5. The microfluidic system of claim 1, wherein there are at least two
sample sources in fluid communication with said sample loading channel on
the same side of said first intersection, and said first load/waste
channel intersects said sample loading channel at a point between said
first intersection and a point at which either of said at least two sample
sources are in fluid communication with said sample loading channel.
6. The microfluidic system of claim 1, wherein said plurality of sample
sources comprises at least four separate sample sources.
7. The microfluidic system of claim 6, wherein at least two separate sample
sources are in fluid communication with said sample loading channel on
each side of said first intersection.
8. The microfluidic system of claim 1, wherein said analysis channel
comprises a buffer reservoir in fluid communication with a first terminus
of said analysis channel, first and second waste reservoirs disposed at
termini of the first and second load/waste channels, respectively, and a
third waste reservoir in fluid communication with a second terminus of
said analysis channel.
9. The microfluidic system of claim 1, wherein said substrate comprises
silica.
10. The microfluidic system of claim 1, wherein said substrate comprises:
a first planar member having a first surface, wherein each of said analysis
channel, sample loading channel and first and second load waste channels
comprise grooves disposed in said first surface; and
a second planar member overlaying and sealably covering said first surface.
11. The microfluidic system of claim 10, wherein each of said plurality of
sample sources comprises a separate hole disposed through said second
planar member, and in fluid communication with a separate sample channel
when said second planar member is overlaying said first planar member,
said separate sample channel being in fluid communication with said sample
loading channel.
12. The microfluidic system of claim 1, wherein each of said separate
sample sources comprises a sample fluid reservoir disposed in said
substrate and in fluid communication with said sample loading channel.
13. The microfluidic system of claim 1, wherein said analysis channel
comprises a separation medium disposed therein.
14. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and in fluid
communication and crossing said analysis channel at a first intersection;
at least six separate sample sources in fluid communication with said
sample loading channel, whereby there is at least one of said at least six
sample sources in fluid communication with said sample loading channel on
each side of said first intersection; and
first and second load/waste channels disposed in said substrate, each of
said first and second load/waste channels intersecting said sample loading
channel at second and third intersections, respectively, said second and
third intersections being on different sides of said first intersection.
15. The microfluidic system of claim 14, wherein at least three separate
sample sources are in fluid communication with said sample loading channel
on each side of said first intersection.
16. The microfluidic system of claim 14, comprising at least eight separate
sample sources.
17. The microfluidic system of claim 16, wherein at least four separate
sample sources are in fluid communication with said sample loading channel
on each side of said first intersection.
18. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and in fluid
communication and crossing said analysis channel at a first intersection;
a plurality of separate sample fluid reservoirs disposed in said substrate
and in fluid communication with said sample loading channel, whereby there
is at least one of said plurality of sample sources in fluid communication
with said sample loading channel on each side of said first intersection,
and wherein a distance from each of said sample fluid reservoirs to said
first intersection via said loading channel is less than two centimeters;
and
first and second load/waste channels disposed in said substrate, each of
said first and second load/waste channels intersecting said sample loading
channel at second and third intersections, respectively, said second and
third intersections being on different sides of said first intersection.
19. The microfluidic system of claim 18, wherein a distance from each of
said plurality of sample fluid reservoirs to said first intersection via
said loading channel is less than one centimeter.
20. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and in fluid
communication and crossing said analysis channel at a first intersection;
a plurality of separate sample sources in fluid communication with said
sample loading channel, whereby there is at least one of said plurality of
sample sources in fluid communication with said sample loading channel on
each side of said first intersection; and
first and second load/waste channels disposed in said substrate, each of
said first and second load/waste channels intersecting said sample loading
channel at second and third intersections, respectively, said second and
third intersections being on different sides of said first intersection;
wherein said analysis channel comprises a buffer reservoir in fluid
communication with a first terminus of said analysis channel, first and
second waste reservoirs disposed at termini of the first and second
load/waste channels, respectively, and a third waste reservoir in fluid
communication with a second terminus of said analysis channel; and
a material direction system for transporting a sample material from each of
said plurality of sample through said sample loading channel and into one
of said first and second waste reservoirs.
21. The microfluidic system of claim 20, wherein said material direction
system comprises:
a different electrode placed in electrical contact with each of said
plurality of different sample sources, said first, second and third waste
reservoirs and said buffer reservoir; and
a voltage source for applying a voltage at each of said different
electrodes.
22. A microfluidic system, comprising:
a planar substrate;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and in fluid
communication and crossing said analysis channel at a first intersection;
a plurality of separate sample sources in fluid communication with said
sample loading channel, whereby there is at least one of said plurality of
sample sources in fluid communication with said sample loading channel on
each side of said first intersection; and
first and second load/waste channels disposed in said substrate, each of
said first and second load/waste channels intersecting said sample loading
channel at second and third intersections, respectively, said second and
third intersections being on different sides of said first intersection;
and
a material direction system for transporting sample material from each of
said plurality of sample sources and injecting at least a portion of said
sample material into said analysis channel.
23. The microfluidic system of claim 22, wherein the substrate comprises
silica.
24. The microfluidic system of claim 22, wherein the substrate comprises a
polymer.
25. The microfluidic system of claim 24, wherein the substrate comprises a
polymer selected from polydimethylsiloxane, polymethylmethacrylate,
polyurethane, polyvinylchloride, polystyrene, polysulfone, and
polycarbonate.
26. The microfluidic system of claim 25, wherein the substrate comprises
polymethylmethacrylate.
27. The microfluidic system of claim 22, wherein the analysis channel
comprises a sieving matrix disposed therein.
28. The microfluidic system of claim 27, wherein the sieving matrix
comprises a linear polyacrylamide polymer.
29. The microfluidic system of claim 28, wherein the linear polyacrylamide
polymer comprises a charged polymer.
30. The microfluidic system of claim 22, wherein the plurality of sample
sources comprises a plurality of separate sample fluid reservoirs disposed
in the substrate.
31. The microfluidic system of claim 22, wherein each of the analysis
channel, sample loading channel and load/waste channels comprises at least
one cross-sectional dimension between about 1 and 100 .mu.m.
32. The microfluidic system of claim 22, wherein the plurality of sample
sources comprises at least four separate sample sources.
33. The microfluidic system of claim 22, wherein the plurality of sample
sources comprises at least eight separate sample sources.
34. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said surface on a first side of said
analysis channel, and intersecting said analysis channel at a first
intersection;
a plurality of sample reservoirs in fluid communication with said sample
loading channel on said first side of said analysis channel;
a waste channel disposed in said substrate on a second side of said
analysis channel, and intersecting said analysis channel at a second
intersection; and
a waste reservoir in fluid communication with said waste channel on said
second side of said analysis channel; and
wherein said plurality of sample reservoirs comprises at least four
separate sample reservoirs.
35. The microfluidic system of claim 34, wherein said plurality of sample
sources comprises at least six separate sample reservoirs.
36. The microfluidic system of claim 34, wherein said plurality of sample
sources comprises at least eight separate sample reservoirs.
37. The microfluidic system of claim 34, wherein the substrate comprises
silica.
38. The microfluidic system of claim 34, wherein the substrate comprises a
polymer.
39. The microfluidic system of claim 38, wherein the substrate comprises a
polymer selected from polydimethylsiloxane, polymethylmethacrylate,
polyurethane, polyvinylchloride, polystyrene, polysulfone, and
polycarbonate.
40. The microfluidic system of claim 39, wherein the substrate comprises
polymethylmethacrylate.
41. The microfluidic system of claim 34, wherein the analysis channel
comprises a sieving matrix disposed therein.
42. The microfluidic system of claim 41, wherein the sieving matrix
comprises a linear polyacrylamide polymer.
43. The microfluidic system of claim 42, wherein the linear polyacrylamide
polymer comprises a charged polymer.
44. The microfluidic system of claim 34, wherein each of the analysis
channel, sample loading channel and load/waste channels comprises at least
one cross-sectional dimension between about 1 and 100 .mu.m.
45. The microfluidic system of claim 34, wherein the plurality of sample
reservoirs comprises at least eight separate sample reservoirs.
46. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
first and second transverse channels, said first transverse channel being
disposed in said substrate on a first side of said analysis channel, and
intersecting said analysis channel at a first intersection, and said
second transverse channel being disposed in said substrate on a second
side of said analysis channel, and intersecting said analysis channel at a
second intersection;
a first sample source in fluid communication with said first transverse
channel;
at least a second sample source in fluid communication with said second
transverse channel;
a first waste channel intersecting said first transverse channel at a third
intersection;
at least a second waste channel intersecting said second transverse channel
at a fourth intersection; and
a material direction system for individually transporting a sample from
each of said first and second sample sources to said first and second
waste channels via said first and second transverse channels,
respectively, and selectively injecting said samples into said analysis
channel.
47. The microfluidic system of claim 46, wherein said first and second
intersections are located at the same point along said analysis channel.
48. The microfluidic system of claim 46, wherein the substrate comprises
silica.
49. The microfluidic system of claim 46, wherein the substrate comprises a
polymer.
50. The microfluidic system of claim 49, wherein the substrate comprises a
polymer selected from polydimethylsiloxane, polymethylmethacrylate,
polyurethane, polyvinylchloride, polystyrene, polysulfone, and
polycarbonate.
51. The microfluidic system of claim 50, wherein the substrate comprises
polymethylmethacrylate.
52. The microfluidic system of claim 46, wherein the analysis channel
comprises a sieving matrix disposed therein.
53. The microfluidic system of claim 52, wherein the sieving matrix
comprises a linear polyacrylamide polymer.
54. The microfluidic system of claim 53, wherein the linear polyacrylamide
polymer comprises a charged polymer.
55. The microfluidic system of claim 46, further comprising at least a
third sample source in fluid communication with the first transverse
channel.
56. The microfluidic system of claim 55, further comprising at least a
fourth sample source in fluid communication with the second transverse
channel.
57. The microfluidic system of claim 56, wherein each of the analysis
channel, first and second transverse channels comprises at least one
cross-sectional dimension between about 1 and 100 .mu.m.
58. A microfluidic system, comprising:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
first and second transverse channels, said first transverse channel being
disposed in said substrate on a first side of said analysis channel, and
intersecting said analysis channel at a first intersection, and said
second transverse channel being disposed in said surface on a second side
of said analysis channel, and intersecting said analysis channel at a
second intersection;
a plurality of sample sources in fluid communication with said first
transverse channel;
a first waste channel intersecting said first transverse channel at a third
intersection;
at least a second waste channel intersecting said second transverse channel
at a fourth intersection; and
a material direction system for individually transporting a sample from
each of said plurality of sample sources to said first and second waste
channels via said first and second transverse channels, respectively, and
selectively injecting said samples into said analysis channel.
59. The microfluidic system of claim 58, wherein said first and second
intersections are located at the same point along said analysis channel.
60. The microfluidic system of claim 58, wherein the substrate comprises
silica.
61. The microfluidic system of claim 58, wherein the substrate comprises a
polymer.
62. The microfluidic system of claim 61, wherein the substrate comprises a
polymer selected from polydimethylsiloxane, polymethylmethacrylate,
polyurethane, polyvinylchloride, polystyrene, polysulfone, and
polycarbonate.
63. The microfluidic system of claim 62, wherein the substrate comprises
polymethylmethacrylate.
64. The microfluidic system of claim 58, wherein the analysis channel
comprises a sieving matrix disposed therein.
65. The microfluidic system of claim 64, wherein the sieving matrix
comprises a linear polyacrylamide polymer.
66. The microfluidic system of claim 65, wherein the linear polyacrylamide
polymer comprises a charged polymer.
67. The microfluidic system of claim 58, wherein the plurality of sample
sources in fluid communication with the first transverse channel comprises
at least four separate sample sources.
68. The microfluidic system of claim 58, wherein the plurality of sample
sources in fluid communication with the first transverse channel comprises
at least six separate sample sources.
69. The microfluidic system of claim 66, wherein each of the analysis
channel, first and second transverse channels comprises at least one
cross-sectional dimension between about 1 and 100 .mu.m.
70. A method of analyzing a plurality of different sample materials,
comprising:
providing a microfluidic device which comprises:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and intersecting said
analysis channel at a first intersection, wherein said sample loading
channel crosses said analysis channel; and
at least first and second sample sources in fluid communication with said
sample loading channel;
a waste reservoir in fluid communication on a same side of said first
intersection as said second sample source;
transporting a first sample from a first of said plurality of sample
sources, through said sample loading channel to said first intersection;
injecting a portion of said first sample into said analysis channel;
analyzing said first sample in said analysis channel;
transporting said second sample through said sample loading channel and
into said waste reservoir; and
injecting a portion of said second sample into said analysis channel.
71. The method of claim 70, wherein said step of transporting said second
sample through said sample loading channel and into said waste reservoir
is carried out substantially concurrently with said step of analyzing said
first sample.
72. The method of claim 70, wherein each of said first and second samples
comprises a plurality of nucleic acid fragments and said analysis channel
comprises a sieving matrix.
73. The method of claim 70, wherein the step of transporting the first
sample comprises electrokinetically transporting the first sample from the
first sample source, through the sample loading channel to the
intersection.
74. The method of claim 70, wherein the step of analyzing the first sample
comprises separating the first sample into constituent elements and
detecting the constituent elements.
75. The method of claim 74, wherein the first sample comprises proteins.
76. A method of performing analysis on a plurality of different sample
materials, comprising:
providing a microfluidic device which comprises:
a planar substrate having a first surface;
an analysis channel disposed in said substrate;
a sample loading channel disposed in said substrate and intersecting said
analysis channel at a first intersection; and
a sample preloading module which comprises at least first and second sample
reservoirs and a waste reservoir, wherein each of said plurality of sample
reservoirs and said waste reservoir are in fluid communication with said
sample loading channel;
transporting a first sample from said first sample reservoir to said first
intersection;
injecting a portion of said first sample into said analysis channel;
concurrently analyzing said portion of said first sample in said analysis
channel and transporting a second sample from said second sample reservoir
into said loading channel and then to said waste reservoir;
transporting said second sample from said loading channel to said
intersection;
injecting a portion of said second sample into said analysis channel; and
analyzing said portion of said second sample in said analysis channel.
77. The method of claim 76, wherein the step of transporting the first
sample comprises electrokinetically transporting the first sample from the
first sample source, through the sample loading channel to the
intersection.
78. The method of claim 76, wherein the step of analyzing the first sample
comprises separating the first sample into constituent elements and
detecting the constituent elements.
79. The method of claim 78, wherein the first sample comprises proteins.
80. The method of claim 76, wherein the sample preloading module further
comprises at least a third sample reservoir in fluid communication with
the sample loading channel, and further comprising the step of
transporting a third sample from said third sample reservoir into said
loading channel and then to said waste reservoir, concurrently with the
step of analyzing said portion of said second sample in said analysis
channel.
81. The method of claim 80, wherein the sample preloading module further
comprises at least a fourth sample reservoir in fluid communication with
the sample loading channel, and further comprising:
injecting a portion of the third sample into said analysis channel;
analyzing the portion of the third sample in the analysis channel; and
transporting a fourth sample from said fourth sample reservoir into said
loading channel and then to said waste reservoir, concurrently with the
step of analyzing said portion of said third sample in said analysis
channel. |
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Claims  |
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Description  |
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BACKGROUND OF THE INVENTION
There has been a growing interest in the development and manufacturing of
microscale fluid systems for the acquisition of chemical and biochemical
information, in both preparative and analytical capacities. Adaptation of
technologies from the electronics industry, such as photolithography, wet
chemical etching and the like, to these fluidic systems has helped to fuel
this growing interest.
One of the first areas in which microscale fluid systems have been used for
chemical or biochemical analysis has been in the area of capillary
electrophoresis (CE). CE systems generally employ fused silica
capillaries, or more recently, etched channels in planar silica
substrates, filled with an appropriate separation matrix or medium. A
sample fluid that is to be analyzed is injected at one end of the
capillary or channel. Application of a voltage across the capillary then
permits the electrophoretic migration of the species within the sample.
Differential electrophoretic mobilities of the constituent elements of a
sample fluid, e.g., due to their differential net charge or size, permits
their separation, identification and analysis. For a general discussion of
CE methods, see, e.g., U.S. Pat. No. 5,015,350, to Wiktorowicz, and U.S.
Pat. No. 5,192,405 to Petersen et al.
Fabrication of CE systems using planar chip technology has also been
discussed. See, e.g., Mathies et al., Proc. Nat'l Acad. Sci. (1994)
91:11348-11352, Jacobsen et al., Anal. Chem. (1994) 66:1114-1118,
Effenhauser et al., Anal. Chem. (1994) 66:2949-2953. However, typically,
such systems employ a single sample introduction point, e.g., a single
well for introducing samples that are to be analyzed in the capillary
channel. This requires rinsing and reloading the well prior to each
analysis. Further, where one wishes to analyze larger numbers of samples,
larger components of each sample, e.g., large nucleic acid fragments,
proteins and the like, can build up within the sample loading and
separation channels, and/or adsorb to capillary walls, eventually
affecting the operation of the system.
It would therefore be desirable to provide microfluidic devices, including
CE systems, which permit faster analysis of multiple samples, and do so
with minimal and even reduced cost, space and time requirements. The
present invention meets these and other needs.
SUMMARY OF THE INVENTION
In a first aspect, the present invention provides a microfluidic system,
that comprises a planar substrate having a first surface. An analysis
channel and a sample loading channel are disposed in the first surface,
whereby the loading channel is in fluid communication and crosses the
analysis channel at a first intersection. A plurality of sample sources
are also provided in fluid communication with the sample loading channel,
whereby at least two of these sample sources are in fluid communication
with the loading channel on different sides of the first intersection.
First and second waste channels intersect the loading channel at second
and third intersections, on different sides of the first intersection. The
system also comprises a material direction system for transporting samples
from each of the sample sources to the loading channel, and for
selectively injecting the samples into the analysis channel.
In a related aspect, the present invention provides a microfluidic system
as described above, but comprising a preloading module, which comprises a
plurality of sample reservoirs and a waste reservoir, wherein each of the
sample reservoirs and the waste reservoir are in fluid communication with
the sample loading channel.
The present invention also provides methods for electrophoretically
analyzing a sample using the devices and systems described herein, which
methods comprise transporting a first sample from the first sample source
through the first sample loading channel to the first intersection. A
portion of the first sample is then injected into the analysis channel and
electrophoreses along the analysis channel. A second sample is then
transported from a second sample source through the loading channel to the
intersection, whereupon a portion of the second sample is injected into
the analysis channel.
In a related aspect, the present invention also provides methods of
electrophoretically analyzing a sample, as described above, and
incorporating a preloading step. The preloading step is carried out
concurrently with the electrophoretic analysis of a first sample. In
particular, the second or subsequent sample is transported from the second
sample reservoir into the loading channel and then to the waste reservoir.
This sample is then injected into the analysis channel as desired.
BRIEF DESCRIPTION OF THE FIGURES
FIGS. 1A-1I schematically illustrates the channel reservoir geometries
employed in the devices of the present invention, and their operation in
loading and injection of multiple samples (FIGS. 1A through 1E) and in
sample preloading (FIGS. 1F through 1I).
FIG. 2 is a schematic illustration of the chronology of the various
material transport steps involved in performing capillary electrophoresis
in a microfluidic device of the present invention (bottom) as compared to
prior art CE systems lacking a preloading feature (top).
FIG. 3 illustrates one embodiment of a microfluidic device incorporating an
improved channel/sample well geometry for performing serial analysis of
multiple samples.
FIG. 4 illustrates another embodiment of a microfluidic device
incorporating an improved channel/sample well geometry for performing
serial analysis of multiple samples.
FIG. 5 is a plot of retention times for fluorescently dyed nucleic acid
fragments injected into a CE channel fabricated into a substrate employing
the improved channel/sample well geometry of the present invention.
FIGS. 6A-6C are plots of fluorescence vs. time for a set of PCR fragments
intercalated with a fluorescent dye (FIG. 6A), PhiX174 DNA, cleaved with
HaeIII and intercalated with a fluorescent dye (FIG. 6B) and a buffer
blank, serially injected into the analysis channel of a microfluidic
device incorporating the channel/sample well geometry of the present
invention.
DETAILED DESCRIPTION OF THE INVENTION
I. General
The present invention generally provides microfluidic devices which
incorporate improved channel and reservoir geometries, as well as methods
of using these devices in the analysis, preparation, or other manipulation
of fluid borne materials, to achieve higher throughputs of such materials
through these devices, with lower cost, material and/or space
requirements.
As used herein, the term "microfluidic device or system" generally refers
to a device or system which incorporates at least two intersecting
channels or fluid conduits, where at least one of the channels has at
least one cross sectional dimension in the range of from | | |