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mutaFISH? (mutation-specific Fluorescence In Situ Hybridization)

Single Cell, Single Molecule, DNA and RNA Mutation Detection at Single Nucleotide Resolution
 
The ability to interrogate cellular heterogeneity at the single cell and single molecule levels has gained increased importance. Many bioreagents and techniques exist to address this issue separately. None has the unified approach to detect and quantify DNA and RNA changes across diverse spectrum particularly point mutations and single-nucleotide polymorphisms (SNP) while maintaining their spatial context. This challenge is even greater as different analytical platforms such fluorescence, cyto/histochemistry, flow cytometry, and microfluidics are needed to study the complexity of biological states.

Abnova has integrated padlock probe and rolling circle amplification (RCA) for in situ, single cell, single molecule, DNA and RNA mutation detection at single nucleotide resolution, and obviated the technical challenges of PCR and hybridization optimization required for efficient and target-specific analysis. Abnova provides a growing portfolio of off-the-shelf, validated mutaFISH? probes and accessory reagents to address the unmet needs in the research and clinical settings.

 
mutaFISH? Workflow

 
Advantage

  • Single Cell, Single Molecule Detection
  • In Situ Analysis of DNA & RNA
  • Single Nucleotide Resolution
  • Higher Sensitivity than dPCR and NGS
  • Multiplexing Capability
  • Cross Analytical Platforms
  • No DNA or RNA Extraction
 
Applications

 
Cell In Situ Hybridization
 
Circulating Tumor Cell In Situ Hybridization
 
Fresh Frozen Tissue In Situ Hybridization
 
Formalin-Fixed Paraffin-Embedded Tissue In Situ Hybridization
Flow Cytometry Mutation-Based Cell Sorting

Microfluidic Cell Isolation & In Situ Hybridization

 
mutaFISH? Probes vs Conventional DNA and RNA FISH Probes

  mutaFISH? Probe Conventional DNA FISH Probe Conventional RNA FISH Probe
Point Mutation Yes No No
Gene Amplification Yes Yes No
Gene Deletion Yes Yes No
Gene Translocation Yes Yes No
Gene Expression Yes No Yes
 
Publication Reference

In situ mutation detection and visualization of intratumor heterogeneity for cancer research and diagnostics.
Grundberg I, Kiflemariam S, Mignardi M, et al. Oncotarget. 2013;4(12):2407-18.
 
In situ detection and genotyping of individual mRNA molecules.
Larsson C, Grundberg I, S?derberg O, Nilsson M. Nat Methods. 2010;7(5):395-7.
 
Lock and roll: single-molecule genotyping in situ using padlock probes and rolling-circle amplification.
Nilsson M. Histochem Cell Biol. 2006;126(2):159-64.
 
In situ genotyping individual DNA molecules by target-primed rolling-circle amplification of padlock probes.
Larsson C, Koch J, Nygren A, et al. Nat Methods. 2004;1(3):227-32.
 
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