Methods for photolithographic synthesis of polymer arrays utilizing anti-reflective coatings
View Patent ↗The present invention provides novel processes for the large scale preparation of arrays of polymer sequences wherein each array includes a plurality of different, positionally distinct polymer sequences having known monomer sequences. The methods of the invention combine high throughput process steps with high resolution photolithographic techniques in the manufacture of polymer arrays.
1. A method of synthesizing an array of polymers, the method comprising:
(a) providing a substrate, wherein the substrate includes an array synthesis surface, and wherein the array synthesis surface comprises functional groups protected by photolabile protecting groups;
(b) deprotecting the functional groups within selected regions of the array synthesis surface with illumination configured to remove the photolabile protecting groups and produce exposed functional groups, wherein the illumination occurs within a synthesis system, wherein the synthesis system comprises a light source and an anti-reflective coating, and wherein the anti-reflective coating reduces reflection of one or more wavelengths of the illumination within the synthesis system;
(c) coupling monomers to the exposed functional groups within the selected regions, wherein the monomers are protected by photolabile protecting groups; and
(d) repeating steps (b)-(c) for one or more repetitions with different sets of selected regions to synthesize the array of polymers.
2. The method of claim 1 , wherein the synthesis system includes a flow cell, wherein the flow cell comprises a flow cell body, wherein the flow cell body and the substrate form a flow cell cavity, and wherein the array synthesis surface is located within the flow cell cavity.
3. The method of claim 2 , wherein the flow cell cavity includes a back surface, wherein the back surface is located opposite the array synthesis surface within the flow cell cavity, and wherein the anti-reflective coating is located on the back surface and reduces reflection of one or more wavelengths off of the back surface.
4. The method of claim 2 , wherein the light source illuminates the selected regions from a side of the substrate which is opposite the array synthesis surface such that the illumination passes through the substrate before reaching the array synthesis surface.
5. The method of claim 2 , wherein step (b) additionally comprises:
adding an index matching fluid to the flow cell cavity before illuminating the selected regions, wherein the index matching fluid possesses an index matching fluid refractive index, wherein the substrate possesses a substrate refractive index, and wherein the index matching fluid refractive index is substantially equivalent to the substrate refractive index.
6. The method of claim 5 , wherein the index matching fluid refractive index is within about 10% of the substrate refractive index.
7. The method of claim 1 , wherein the substrate is a slide.
8. The method of claim 1 , wherein the substrate comprises a material selected from the group consisting of glass, quartz and silicon.
9. The method of claim 1 , wherein the illumination includes one or more wavelengths longer than 340 nm.
10. The method of claim 9 , wherein the illumination includes a wavelength of 365 nm.