Plastic microfluidic separation and detection platforms
Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.
1. A plastic chip component, comprising:
(a) a separation chip having a substrate layer and a cover layer;
wherein one of the substrate layer and cover layer includes a plurality of grooves and the other of the substrate layer or the cover layer includes a series of through holes;
wherein a top surface of the substrate layer and a bottom surface of the cover layer are bonded together to form (i) a first microfluidic channel that connects the sample to one of a plurality of second microfluidic channels; and (ii) each of said plurality of second microfluidic channels having a detection region at the termini of a separation region;
wherein each of the substrate layer and the cover layer comprises a thin plastic of polyethylene, a poly(carbonate), an unsaturated, partially unsaturated or saturated cyclic olefin polymer (COP), an unsaturated, partially unsaturated, or saturated cyclic olefin copolymer (COC), or a norbornene thermopolymer, or a poly (acrylate) which essentially does not fluoresce light having a wavelength between 500 and 800 when excited at a wavelength between about 450 and 500 nm,
and wherein each of the substrate layer and the cover layer independently has a thickness of less than 200 μm; and
(b) a plastic support fixedly attached to the chip having
an anode portion with at least one anode well,
a cathode portion with at least one cathode well and
a center portion therebetween, the center portion having a detection window fabricated from a cut-out in the plastic support which overlaps with the detection region of each of the plurality of second microfluidic channels such that the total thickness of plastic at the detection region is less than 400 μm.
2. The chip of claim 1 , wherein the thin plastic is a norbornene thermopolymer.
3. The chip of claim 1 , wherein the plastic support comprises polyethylene, a poly(acrylate), a poly(carbonate), an unsaturated, partially unsaturated or saturated cyclic olefin polymer (COP), an unsaturated, partially unsaturated, or saturated cyclic olefin copolymer (COC), or a norbornene thermopolymer.
4. The chip of claim 3 , wherein the plastic support is made from an acrylic sheet.
5. The chip of claim 1 , wherein the detection region of each of the plurality of second microfluidic channels is in substantially the same location along each channel such that the detection window is in a single location in the center portion of the support.
6. The chip of claim 1 , wherein each of the plurality of second microfluidic channels has a separation length of from 2 cm to 50 cm.
7. The chip of claim 1 , wherein said first microfluidic channel further comprises an injection channel.
8. The chip of claim 3 , wherein the thin plastic essentially does not fluoresce light having a wavelength at 570 nm.
9. The chip of claim 1 enabling a user to detect a plurality of nucleic acid species in a nucleic acid sample generated for fragment sizing applications with a signal to noise of greater than 3 starting with a single copy of a nucleic acid template for PCR amplification.
10. The chip of claim 1 enabling a user to detect a plurality of nucleic acid species in a nucleic acid sample generated for DNA sequencing application with a signal to noise of greater than 3 starting with a single copy of a DNA template for PCR amplification.
11. The chip of claim 1 , wherein each of the plurality of first or second microfluidic channels further comprises a surface coating, preferably wherein the surface coating is hydroxypropylmethylcellulose (HPMA), poly(ethylene oxide) (PEO), poly(vinyl alcohol)(PVA), poly(dimethyl acrylamide) (PDMA), poly(vinylpyrrolidinone), dimethylacrylamide(DMA), diethylacrylamide DEA, poly(diethylacrylamide) and mixtures thereof.
12. The chip of claim 1 , wherein the each of the plurality of second microfluidic channels further comprises a sieving matrix, preferably wherein the sieving matrix comprises a linear or cross-linked poly (N,N-dialkylacrylamide), linear polyacrylamide, polydimethylacrylamide, polyvinylpyrrolydinone, or combinations thereof, more preferably wherein the sieving matrix comprises 1-50 wt. % polyacrylamide.
13. The chip of claim 1 , further comprising a porous layer between each of said at least one cathode wells and each of said first or second microfluidic channels wherein the porous layer is capable of substantially blocking passage of gas bubbles from each of said at least one cathode wells into each of said first or second microfluidic channels.
14. The chip of claim 13 , wherein the porous layer comprises a glass frit, a polymer frit, a polymer membrane, or a polymer filter.
15. The chip of claim 1 wherein said first microfluidic channel contains an amplification product.
16. The chip of claim 1 wherein said first microfluidic channel contains product from a sequencing reaction.
17. The chip of claim 1 wherein said first microfluidic channel contains amplified human genomic DNA and sizing standard.
18. The chip of claim 15 wherein the amplification product contains amplified equine DNA for racehorse breeding and tracking.
19. The chip of claim 15 wherein the amplification product contains animal DNA for livestock breeding.
20. The chip of claim 15 wherein the amplification product contains animal DNA for pet identification.
21. The chip of claim 15 wherein the amplification product contains amplified biological weapons agents.