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US 8,617,895 B2Granted
Polycationic quaternary ammonium polymer coatings for immobilizing biological samples
AssigneeTriPath Imaging, Inc.
InventorsWilliam Alan Fox, William Carl Ray, III
Life of this patent
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Abstract
The present invention is directed to a pre-coated substrate, such as a slide, that is useful for immobilizing a sample. The invention is further provides methods of preparing such pre-coated substrates and methods of analyzing biological samples immobilized on such pre-coated substrate. The substrate is coated with a polycationic polymeric coating material specifically selected such that that coated substrate exhibits increased stability and prolonged shelf-life. Preferred polymeric coating materials include allylic or vinylic polymers having cationic groups thereon and having no more than a small percentage of peptidic monomeric linkages, particularly polydiallyldimethylammonium (PDDA).
Claims41
A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:
- providing a glass substrate having a surface comprising a plurality of anionic groups;
- contacting the substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material comprising at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof, said quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of a single layer of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the substrate with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material;
- wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da, and
- drying the coating of the non-peptidic, quaternary ammonium polymeric material.
1 more element
A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:
- providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups;
- contacting the surface of the microscope slide with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material;
- wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da,
- drying the coating of the non-peptidic, quaternary ammonium polymeric material;
- and
- rinsing the microscope slide having the dried coating of the non-peptidic, quaternary ammonium polymeric material thereon;
- wherein the microscope slide surface has not been subjected to a cleaning process after receipt from the manufacturer and prior to contacting the microscope slide surface with the solution of the non-peptidic, quaternary ammonium polymeric material.
4 more elements
A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:
- providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups;
- and
- contacting the surface of the microscope slide with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.
A method for preparing a microscope slide adapted for immobilizing a biological sample, said method comprising:
- providing a previously manufactured microscope slide having a surface comprising a plurality of anionic groups;
- and
- contacting the surface of the microscope slide with a composition comprising a solution of polydiallyldimethylammonium present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the polydiallyldimethylammonium on at least a portion of the surface of the microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the polydiallyldimethylammonium and with no further layers of any material coated over said layer of the polydiallyldimethylammonium, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.
A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:
- providing a glass substrate having a surface comprising a plurality of anionic groups;
- and
- contacting the surface of the glass substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material present in a concentration of about 0.01% (w/v) to about 10.0% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the glass substrate microscope slide with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric material, and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material, wherein the non-peptidic, quaternary ammonium polymeric coating material has an average molecular weight of about 400,000 Da to about 500,000 Da.
A method for preparing a coated substrate adapted for immobilizing a biological sample, said method comprising:
- providing a glass substrate having a surface comprising a plurality of anionic groups;
- contacting the substrate with a composition comprising a solution of a non-peptidic, quaternary ammonium polymeric material comprising at least one monomer selected from the group consisting of diallyldimethylammonium, methylacrylamidopropyltrimethylammonium, methacryloyloxyethyltrimethylammonium, 4-vinyl-benzyltrimethylammonium, acryloxyethyldimethylbenzyl ammonium, acryloxyethyltrimethyl ammonium, methacryloxyethyldimethylammonium, methacryloxyethyltrimethylbenzylammonium, trimethyl-2-methacryloylethylammonium, trimethyl-2-methacrylaminopropylammonium, and mixtures thereof, said quaternary ammonium polymeric material present in a concentration of about 0.15% (w/v) to about 0.75% (w/v) in solution, the solution having a pH of about 8 to about 14, to form a coating of a single layer of the non-peptidic, quaternary ammonium polymeric material on at least a portion of the surface of the substrate with no intervening layers of a different coating material sandwiched between two or more layers of the non-peptidic, quaternary ammonium polymeric coating material and with no further layers of any material coated over said layer of the non-peptidic, quaternary ammonium polymeric material and
- drying the coating of the non-peptidic, quaternary ammonium polymeric material.
Dependencies are inferred from the claim text.