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. 2016 Sep 5;4:89. doi: 10.3389/fcell.2016.00089

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Copyright © 2016 Cui, Shu and Li.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Single molecule FISH techniques for research application in single cells. Schematic drawings of single molecule FISH methods: (A) Cas-9 mediated FISH (CASFISH) using fluorophore-coupled sgRNAs and dCAS9. (B) Oligopaint-FISH using fluorophore-coupled primary oligonucleotides for targeted SNP loci and fluorophore-coupled second oligonucleotide to enhance labeling efficiency shows differential labeling of paternal (pat) and maternal (mat) chromosomes. (C) single molecule RNA FISH by rolling cycling amplification (RCA) using padlock probes targeting to reverse transcripted cDNA with different alleles followed by ligation, cycling amplification and specific fluorophore-couple probe hybridization and visualization. (D) Sequential barcoding of multiplex different mRNAs by repeat rounds of hybridization, imaging, and stripping. Star, diamond, and triangle are symbols for different fluorophores.