Method of preparing fluorescently labeled microbeads

Claim 1

A method for preparation of polymeric microbeads comprising: (a) contacting a dispersion of swelled polymeric particles with an ethylenically unsaturated monomer containing an epoxy group, said monomer being absorbed into said particles; (b) polymerizing said monomer to cause additional swelling of said particles, whereby said swelled particles provide a microbead with an epoxide group to serve as a first reactive functional group; (c) reacting said epoxide with a polyfunctional spacing agent thereby to provide a second reactive functional group; and (d) reacting said second reactive functional group with a suitable reagent which provides said microbead with a detectable label.

Claim 2

A method for preparation of polymeric microbeads comprising: (a) contacting a dispersion of swelled polymeric particles with an ethylenically unsaturated monomer containing an epoxy group, said monomer being absorbed into said particles; (b) polymerizing said monomer to cause additional swelling of said particles, whereby said swelled particles provide a microbead with an epoxide group to serve as a first reactive functional group; (c) reacting said epoxide with a polyfunctional spacing agent thereby to provide a second reactive functional group; and (d) reacting said second reactive functional group with a suitable reagent which provides said microbead with a detectable label.

Claim 3

A method for preparation of polymeric microbeads comprising: (a) contacting a dispersion of polymeric particles with a substance which is absorbed by said particles thereby to cause swelling of said particles; (b) contacting said swelled particles with at least one monomer having the formula: ##STR6## wherein R and R 1 are independently selected from the group consisting of hydrogen or lower alkyl of 1 to 6 carbon atoms and A is selected from the group consisting of a methylene or carbonyl group whereby said monomers are absorbed into said particles thereby to provide said particles with an epoxide group to serve as a first reactive functional group; (c) polymerizing said monomers to cause additional swelling of said particles whereby said swelled particles provide a microbead with an epoxide group to serve as a first reactive functional group; (d) reacting said epoxide group with a polyfunctional spacing agent of the formula: X--Q--Y wherein X is selected from the group consisting of amino, hydroxyl or sulfhydryl, Q is selected from the group consisting of a straight or branched chain alkylene group of 2 to 10 carbon atoms, an aryl group or a heterocyclic group, and Y is selected from the group consisting of amino, hydroxyl, sulfhydryl, halo or carboxyl, thereby to provide said microbead with a second reactive functional group; and (e) reacting said second reactive functional group with a fluorescent reagent selected from the group consisting of a derivative of fluorescein, a derivative of rhodamine or a derivative of phycobiliprotein.

Claim 4

A method for preparation of polymeric microbeads comprising: (a) contacting a dispersion of polymeric particles with a substance which is absorbed by said particles thereby to cause swelling of said particles; (b) contacting said swelled particles with at least one monomer having the formula: ##STR6## wherein R and R 1 are independently selected from the group consisting of hydrogen or lower alkyl of 1 to 6 carbon atoms and A is selected from the group consisting of a methylene or carbonyl group whereby said monomers are absorbed into said particles thereby to provide said particles with an epoxide group to serve as a first reactive functional group; (c) polymerizing said monomers to cause additional swelling of said particles whereby said swelled particles provide a microbead with an epoxide group to serve as a first reactive functional group; (d) reacting said epoxide group with a polyfunctional spacing agent of the formula: X--Q--Y wherein X is selected from the group consisting of amino, hydroxyl or sulfhydryl, Q is selected from the group consisting of a straight or branched chain alkylene group of 2 to 10 carbon atoms, an aryl group or a heterocyclic group, and Y is selected from the group consisting of amino, hydroxyl, sulfhydryl, halo or carboxyl, thereby to provide said microbead with a second reactive functional group; and (e) reacting said second reactive functional group with a fluorescent reagent selected from the group consisting of a derivative of fluorescein, a derivative of rhodamine or a derivative of phycobiliprotein.

Claim 5

A method for the preparation of polymeric microbeads comprising the steps of: (a) providing an emulsion of dichloroethylene, sodium lauryl sulfate and benzoyl peroxide; (b) adding 1-chlorododecane to said emulsion and homogenizing said emulsion; (c) adding acetone and polyvinyl toluene seed particles to said homogenized emulsion to provide a seed particle mixture; (d) removing said acetone; (e) adding to said seed particle mixture a potassium chloride solution and a monomer mixture containing glycidyl methacrylate, methyl methacrylate and sodium dilaurylsulfonate dispersed in a potassium chloride solution; (f) polymerizing said monomer mixture to provide a uniform microbead having an epoxide group; (g) stirring said microbeads with an aqueous solution of 1,3-diaminopropane; and (h) stirring said microbeads with a solution of fluorescein isothiocyanate in dimethylformamide.

Claim 6

A method for the preparation of polymeric microbeads comprising the steps of: (a) providing an emulsion of dichloroethylene, sodium lauryl sulfate and benzoyl peroxide; (b) adding 1-chlorododecane to said emulsion and homogenizing said emulsion; (c) adding acetone and polyvinyl toluene seed particles to said homogenized emulsion to provide a seed particle mixture; (d) removing said acetone; (e) adding to said seed particle mixture a potassium chloride solution and a monomer mixture containing glycidyl methacrylate, methyl methacrylate and sodium dilaurylsulfonate dispersed in a potassium chloride solution; (f) polymerizing said monomer mixture to provide a uniform microbead having an epoxide group; (g) stirring said microbeads with an aqueous solution of 1,3-diaminopropane; and (h) stirring said microbeads with a solution of fluorescein isothiocyanate in dimethylformamide.