Methods of making fluorescent microspheres
First Claim
1. A method of forming a dyed microsphere comprising a first step of forming an acrylic latex bead having on its surface at least five thousand ester terminations, and thereafter a second step of simultaneously reacting said ester terminations on said acrylic latex bead with an amine-terminated dye and with a diamine to form by transacylation a conjugate of said amine-terminated dye and said diamine with said bead.
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Abstract
Highly fluorescent latex microspheres have a diameter of less than five hundred angstroms and have more than five thousand fluorescent markers per sphere. The microspheres are prepared by reacting an acrylic latex bead with a diamine and a fluorescent amine at elevated pH. A protein such as avidin or an immunoglobulin may then be conjugated to the diamine. A single fluorescent microsphere is visible using standard fluorescent microscopy. Therefore the microspheres may be utilized not only to visualize cell surface anitgens but also DNA encoding for single genes, by means of a biotinylated DNA probe.
358 Citations
20 Claims
- 1. A method of forming a dyed microsphere comprising a first step of forming an acrylic latex bead having on its surface at least five thousand ester terminations, and thereafter a second step of simultaneously reacting said ester terminations on said acrylic latex bead with an amine-terminated dye and with a diamine to form by transacylation a conjugate of said amine-terminated dye and said diamine with said bead.
- 17. The method of forming a fluorescent microsphere comprising forming a latex bead, thereafter a step of chemically attaching to said latex bead a first set of spacers having attached thereto fluorescent labels and simultaneously chemically attaching to said latex bead a second set of spacers, and thereafter a step of chemically attaching to said second set of spacers a selectively reactive means for selectively reacting with a chemical species on a substrate to label said substrate.
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