Related Experiment Video
Updated: Sep 11, 2025
10:14
Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase
Published on: November 8, 2019
6.4K
A simple, ultrastable, and cost-effective oxygen-scavenging system for long-term DNA-PAINT imaging
Rebecca T Perelman1,2, George M Church1,3,4, Johannes Stein1,3
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
Biorxiv : the Preprint Server for Biology
|August 12, 2025
Summary
We developed a new, enzyme-free buffer using sodium sulfite and Trolox to prevent damage during DNA-PAINT super-resolution microscopy. This stable, cost-effective solution enhances long-term imaging by protecting DNA structures.
Area of Science:
- Super-resolution microscopy
- Molecular imaging
- Biotechnology
Background:
- DNA-PAINT enables nanoscale imaging via transient DNA binding.
- Photobleaching resistance is high, but reactive oxygen species (ROS) limit long-term imaging.
- Current oxygen-scavenging systems (OSS) degrade, reducing efficacy and robustness.
Purpose of the Study:
- To develop a stable, enzyme-free oxygen-scavenging buffer for extended DNA-PAINT imaging.
- To overcome the limitations of current enzymatic OSS.
- To enhance the robustness, cost-effectiveness, and performance of DNA-PAINT.
Main Methods:
- Formulation of an enzyme-free buffer using sodium sulfite (Na2SO3) and Trolox (SST).
- Evaluation of docking strand integrity and localization sampling over extended imaging periods.
- Comparison of SST performance against existing OSS.
Main Results:
- The SST buffer effectively preserves docking strand integrity for over 24 hours.
- SST demonstrates tenfold improvement in buffer stability compared to enzymatic OSS.
- SST reduces costs by over 90% and is easy to prepare.
Conclusions:
- The enzyme-free SST buffer provides a robust, cost-effective, and high-performance solution for extended DNA-PAINT imaging.
- SST mitigates ROS damage, enhancing localization sampling and buffer stability.
- This new OSS advances the utility of DNA-PAINT for long-term nanoscale investigations.
