Protective surface coatings for flow cells
An example of a method includes providing a substrate with an exposed surface comprising a first chemical group, wherein the providing optionally comprises modifying the exposed surface of the substrate to incorporate the first chemical group; reacting the first chemical group with a first reactive group of a functionalized polymer molecule to form a functionalized polymer coating layer covalently bound to the exposed surface of the substrate; grafting a primer to the functionalized polymer coating layer by reacting the primer with a second reactive group of the functionalized polymer coating layer; and forming a water-soluble protective coating on the primer and the functionalized polymer coating layer. Examples of flow cells incorporating examples of the water-soluble protective coating are also disclosed herein.
1. A flow cell, comprising:
a substrate including a bonding region at its periphery;
a lid bonded to the bonding region of the substrate;
a spacer layer positioned between the substrate and the lid so that the spacer layer contacts the bonding region, bonds the substrate and lid together, and forms sidewalls of the flow cell, wherein the lid and the substrate and the spacer layer define a flow channel between the substrate and the lid, and wherein the lid bonded to the substrate at the bonding region forms a boundary of the flow channel;
an input port and an output port formed in the lid or the substrate;
surface chemistry positioned on at least a portion of a surface of the substrate in the flow channel, the surface chemistry including:
a functionalized polymer coating layer covalently bound to a chemical group on the surface; and
a primer grafted to the functionalized polymer coating layer; and
a water-soluble protective coating applied on the primer and the functionalized polymer coating layer.
2. The flow cell of claim 1 , wherein the substrate is a patterned substrate comprising depressions separated by interstitial regions, and wherein a fluid introduced into the flow channel is introduced into the depressions.
3. The flow cell of claim 2 , wherein the water-soluble protective coating is in the depressions and on at least a portion of the interstitial regions.
4. The flow cell of claim 1 , wherein the substrate is a non-patterned substrate.
5. The flow cell of claim 1 , wherein the water-soluble protective coating comprises: a non-cationic synthetic polymer; a natural polysaccharide or a derivative thereof; a natural protein or a derivative thereof; a water-soluble salt; or a small molecule compound selected from the group consisting of a water-soluble surfactant, a sugar, an antioxidant, a chelator, a buffer, a glycol, glycerol, and a cyclodextrin; or a combination thereof.
6. The flow cell of claim 1 , wherein the water-soluble protective coating comprises a polyvinyl alcohol/polyethylene glycol graft copolymer, sucrose, dextran, a polyacrylamide, a glycol, tris(hydroxymethyl)aminomethane or a salt thereof, ethylenediaminetetraacetic acid or a salt thereof, (tris(2-carboxyethyl)phosphine), tris(3-hydroxypropyltriazolylmethyl)amine or a salt thereof, bathophenanthrolinedisulfonic acid disodium salt, a hydroxyl functional polymer, glycerol, or saline sodium citrate, or a mixture thereof.
7. The flow cell of claim 1 , wherein the functionalized polymer coating layer is poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide (PAZAM).
8. The flow cell of claim 7 , wherein the surface of the substrate is silanized with a silane or silane derivative and wherein PAZAM is covalently attached to the silane or the silane derivative.
9. The flow cell of claim 8 , wherein the silane or silane derivative is [(5-bicyclo[2.2.1]hept-2-enyl)ethyl]trimethoxysilane.
10. A method of using the flow cell of claim 1 , comprising:
inserting the flow cell into a sequencing instrument; and
removing the water-soluble protective coating by exposing the water-soluble protective coating on the flow cell to water.
11. The method of claim 10 , wherein the removing is done by performing a flow through process to remove the water-soluble protective coating.