DNA-PAINT allows for <10 nm localization precision in single molecule localization microscopy
DNA-PAINT is a technique to enable single molecule localization through the binding of short (<10 nucleotides) oligonucleotides labeled with a fluorophore to a complementary oligonucleotide bound to a target molecule, typically an antibody or nanobody. The binding of the short oligos is transient in nature, and thus creates a blinking effect akin to dSTORM or PALM. DNA-PAINT has numerous advantages over other blinking techniques:
- Higher photon yield. The blinks are typically longer lasting than in conventional dSTORM resulting in higher photon yields from the fluorophore, therefore allowing for a much higher localization precision (<10 nm) when compared to such methods as dSTORM and PALM.
- The imaging is practically unbleachable. The sample is bathed in an excess of fluorophore, allowing for extremely long-lasting imaging.
- Unlimited multiplexing potential. Since target specificity is set by nucleotide sequence, multiple targets can be labeled with different oligo sequences. The Vutara’s microfluidics unit can be utilized such that the imaging strand for a given target can be washed from the sample and different imaging strands labeling different biological targets can be added.
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