TECHNOLOGY
WHAT IS DNA-PAINT?
DNA-PAINT (DNA-Point Accumulation for Imaging in Nanoscale Topography) is a localization based super-resolution microscopy method that uses short, dye-labeled single stranded DNA (ssDNA strands) that transiently bind to their complements:
- Imager strand – a dye-labeled DNA oligo present in imaging buffer
- Docking strand – a DNA oligo complementary to the imager strand that is delivered to the target of interest via binders (e.g. antibodies, single domain antibodies)
When a dye-labeled imager transiently hybridizes to a docking strand in DNA-PAINT imaging buffer, fluorescent signal or “blinking” occurs. To create a super-resolution image, a camera captures blinking over a period of time (10 – 50 min) and the resulting image is post-processed using Picasso software.
WHY USE DNA-PAINT?
- Ultra resolution! Resolve 10 – 15 nm distances in cells.
- Unlimited target multiplexing! In DNA-PAINT, spectrally compatible dyes aren’t needed to image multiple targets at a time. Instead, you can simply couple different docking strands (e. different DNA sequences) to your target binders and image each target sequentially using the same laser line with its corresponding imager strand.
- Target Counting! The predictable nature of DNA binding kinetics allows you to quantify how many proteins are present in cells (qPAINT).
WHERE TO START?
You need 3 things to do DNA-PAINT:
- Simple TIRF Microscope – like the ZEISS Elyra 7, Nikon N-STORM, ONI Nanoimager, abbelight SAFe 180, Bruker VUTARA VXL microscope, Andor Dragonfly or similar system found in your imaging facility.
- DNA-PAINT compatible binder – any antibody or single-domain antibody that binds to your target protein and is coupled to a docking strand. Find a DNA-PAINT compatible binder here at Massive Photonics! We have a wide range of antibodies and single domain antibodies. Order your custom antibody or single-domain antibody (PRODUCTS page).
- Picasso data analysis software – a free software developed for DNA-PAINT image analysis (Download Picasso). Find out how to use Picasso in Schnitzbauer, Strauss et al.