Combined with Bruker's software-controlled, automated, fully-integrated Microfluidics Unit, the Vutara VXL super-resolution microscope is the only commercial system that offers out-of-the-box multiplexed DNA-PAINT capabilities. Performed on these tools, DNA-PAINT imaging allows for sub-10 nm localization precision in single-molecule localization microscopy with unlimited multiplexing potential.
Moreover, the Vutara microscope with microfluidics system provides the added benefits of:
Or contact us to learn more about performing DNA-PAINT experiments with the Vutara VXL microscope.
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. It allows for extensive imaging modalities, from whole-cell extensive Z-stacks to high-resolution multi-target images.
DNA-PAINT works through the transient binding of a short “imaging oligonucleotide” containing a fluorophore to a complementary oligonucleotide — called the “docking strand” — on the target of interest, such as an antibody, nanobody, aptamer or suicide enzyme ligand.
Super-resolution imaging with DNA-PAINT involves several steps, including:
Since the sample is bathed in a large excess of constantly exchanging imaging strand, the target is essentially unbleachable, making it possible to batch-process a large number of frames and extended Z-stacks.
DNA-PAINT allows sub-10 nm localization precision, making it one of the most precise microscope techniques available.
Here, a Vutara microscope with a water immersion 1.2 NA objective was utilized for a DNA-PAINT experiment. The image shows a whole BS-C-1 cell’s tubulin network labeled with tubulin antibodies conjugated to a DNA-PAINT secondary antibody. The inset shows a zoomed-in section of the tubulin network. The lumen of the microtubule is clearly visible.
Multiplexed multicolor super-resolution imaging is made possible with DNA-PAINT.
DNA-PAINT has the potential for large-scale multicolor Z-stacks due to the fact the sample is bathed in a practically limitless supply of fluorophore. This enables large-scale z-stack imaging composed of millions of localizations.
Here, a two-color DNA-PAINT experiment was performed on the Vutara single-molecule localization microscope. Tubulin is labeled in cyan and clathrin in magenta. Furthermore, due to the unbleachable nature of DNA-PAINT large Z-stacks are possible.
DNA-PAINT has enormous potential for multiplexed imaging.
Here, a multi-target DNA-PAINT experiment was performed using the Vutara VXL and integrated fluidics unit. Using orthogonal docking strands on different probes, a potentially unlimited number of targets are possible.
Also pictured is a four-target DNA-PAINT experiment performed on the Vutara single-molecule localization microscope and integrated fluidics unit. F-actin-magenta, tom20-cyan, tubulin-yellow and clathrin-green.
BS-C-1 labeled with anti-tubulin, actin, anti-tom20 and anti-clathrin. Orthogonal 2º DNA-PAINT antibodies were purchased from Massive-Photonics.com.