Compositions for multiplex imaging using labeled nucleic acid imaging agents

Abstract

The present application provides compositions for multiplexed imaging using labeled nucleic acid imaging agents.

What is claimed is: 
     
         1 . A composition comprising:
 a sample bound to more than one target-specific binding partners, each binding partner bound to an oligonucleotide sequence and at least one docking strand stably bound to a labeled imager strand, directly or indirectly, wherein the oligonucleotide sequence is a docking strand or a primer strand and if the oligonucleotide sequence is
 (a) a docking strand, more than one docking strands are associated with each target-specific binding partner, or 
 (b) a primer strand, more than one docking strands are associated with the primer strand. 
   
     
     
         2 . The composition of  claim 1 , wherein the docking strand bound to a labeled imager strand, directly or indirectly, comprises at least 90% binding for 30 minutes. 
     
     
         3 . The composition of  claim 1 , wherein the (a) docking strand or (b) docking strand and any intermediate strand is 80 nucleotides or less, 70, 60, 50, 40, or 30 nucleotides or less. 
     
     
         4 . The composition of  claim 1 , wherein the imager strand is 60 nucleotides or less. 
     
     
         5 . A kit for detecting more than one target in a sample, the kit comprising:
 (i) more than one target-specific binding partner-nucleotide conjugates, each comprising a target-specific binding partner linked to a nucleotide sequence, wherein different target-specific binding partners are linked to different nucleotide sequences;   wherein the nucleotide sequence associated with each target-specific binding partner is a docking strand or a primer strand for initiating amplification of a docking strand, and wherein the number of docking strands associated with each target-specific binding partner is capable of being amplified;   (ii) at least one labeled imager strand comprising an imager strand with a label attached thereto,   wherein the docking strand is capable of being stably bound to the at least one labeled imager strand complementary to the docking strand, directly or indirectly;   (iii) reagents for amplification of the number of docking strands, including an enzyme for amplification, optionally an amplifier strand and nucleotides.   
     
     
         6 . The kit of  claim 5 , wherein at least 70% of the labeled imager strands bound to the complementary docking strands, directly or indirectly, remain bound for at least 30 minutes. 
     
     
         7 . The kit of  claim 5 , wherein at least 90% of the labeled imager strands bound to the complementary docking strands, directly or indirectly, remain bound for at least 30 minutes. 
     
     
         8 . The kit of  claim 5 , wherein the labeled imager strands bound to the complementary docking strands, directly or indirectly, remain bound for at least 5 minutes. 
     
     
         9 . The kit of  claim 5 , wherein the labeled imager strands bound to the complementary docking strands, directly or indirectly, remain bound for at least 30 minutes. 
     
     
         10 . The kit of  claim 5 , wherein the docking strand is stably bound to the at least one labeled imager strand indirectly through an intermediate strand. 
     
     
         11 . The kit of  claim 10 , wherein (i) the intermediate strand is stably bound to the docking strand or (ii) a labeled imager strand is stably bound to the intermediate strand or both (i) and (ii) are met. 
     
     
         12 . The kit of  claim 5 , wherein the docking strand is 80 nucleotides or less in length. 
     
     
         13 . The kit of  claim 10 , wherein the docking strand and intermediate strand is 80 nucleotides or less in total length. 
     
     
         14 . The kit of  claim 5 , wherein the imager strand is 60 nucleotides or less in length. 
     
     
         15 . The kit of  claim 5 , wherein the enzyme for amplification is a polymerase. 
     
     
         16 . The kit of  claim 5 , wherein the amplifier strand is included and is linear or nonlinear. 
     
     
         17 . The kit of  claim 16 , wherein the amplifier strand has complementarity to the docking strand and comprises a sequence identical to the imager strand.