Microarray fabrication system and method
A method includes forming a patterned substrate including a plurality of base pads, using a nano-imprint lithography process. A capture substance is attached to each of the plurality of base pads, optionally through a linker, the capture substance being adapted to promote capture of a target molecule.
1. A method comprising:
forming a patterned substrate that comprises an array of polymer base pads at predetermined sites on a substrate, the forming involving:
patterning a photoresist layer on the substrate via lithography to expose interstitial spaces;
applying a passivation layer to the interstitial spaces, the passivation layer being selected from the group consisting of diamond-like carbon, hexa-methyldisilizane, polytetrafluoroethylene, and parylene;
lifting off the photoresist layer to yield a patterned surface with inert pads; and
applying a patterning medium using a microcontact printing process on the inert pads to form the polymer base pads with the passivation layer disposed on the substrate between the polymer base pads, wherein the patterning medium includes a poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide (PAZAM) polymer;
wherein each of the polymer base pads comprises a molecule binding substance disposed over the polymer base pad and configured to promote capture of a target nucleic acid molecule at each of the polymer base pads.
2. The method of claim 1 , wherein the microcontact printing process comprises contacting a stamp that includes a plurality of pillars coated with the patterning medium on substrate, so as to transfer the patterning medium to the inert pads on the substrate and thereby form the polymer base pads on the substrate.
3. The method of claim 1 , wherein the molecule binding substance comprises a primer nucleic acid.
4. A biological microarray system, comprising:
an array of polymer base pads at predetermined sites on a substrate, wherein the polymer base pads include a poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide) polymer;
a passivation layer disposed on the substrate between the polymer base pads, the passivation layer selected from the group consisting of diamond-like carbon, hexa-methyldisilizane, polytetrafluoroethylene, and parylene; and
a molecule binding substance disposed over each of the polymer base pads configured to capture a target nucleic acid molecule at each of the polymer base pads.
5. The biological microarray system of claim 4 , wherein the molecule binding substance comprises a primer nucleic acid.
6. A biological microarray system, comprising:
an array of metal base pads at predetermined sites on a substrate, the metal base pads selected from the group consisting of gold, titanium, nickel, and copper;
a capture substance linked to each of the base pads and configured to capture a single target nucleic acid template molecule at each of the base pads;
a porous attachment layer disposed over the base pads; and
two different primer nucleic acids attached to the porous attachment layer, wherein the two different primer nucleic acids form a pair for amplification of the single target nucleic acid template molecule.
7. The system of claim 6 , wherein each of the two different primer nucleic acids provides an extendable 3′ terminus for copying the target nucleic acid molecule.
8. The system of claim 6 , wherein the substrate is glass.
9. The system of claim 6 , wherein the porous attachment layer is patterned and defines a plurality of regions at least partially surrounding each of the base pads.
10. The system of claim 6 , wherein the porous attachment layer extends substantially continuously over the plurality of base pads.
11. The system of claim 6 , wherein the porous attachment layer comprises a hydrogel, a polyacrylamide copolymer, or poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide).
12. The system of claim 6 , wherein the capture substance is selected from the group consisting of a silane, a biotin-binding protein, avidin, streptavidin, and neutravidin.
13. The system of claim 6 , wherein a size of each base pad ranges from about 30 nm to about 500 nm.
14. A biological microarray system, comprising:
an array of metal base pads at predetermined sites on a substrate, wherein each base pad is a nanodot or a nanoparticle and each nanodot or nanoparticle is selected from the group consisting of gold, titanium, nickel, and copper;
a molecule binding substance attached to each base pad and configured to capture a target nucleic acid molecule at each of the base pads;
a porous attachment layer disposed over the base pads; and
primer nucleic acids attached to the porous attachment layer.
15. A biological microarray system, comprising:
an array of metal base pads at predetermined sites on a substrate, wherein the metal base pads are gold base pads;
a capture substance linked to each of the base pads;
a molecule binding substance attached to the capture substance and configured to capture a target nucleic acid molecule at each of the base pads;
wherein the capture substance with the molecular binding substance attached is thiol-avidin;
a porous attachment layer disposed over the base pads; and
primer nucleic acids attached to the porous attachment layer.
16. The system of claim 15 , wherein the target nucleic acid molecule is a template DNA molecule containing biotin on one end.