Good news for researchers using high-resolution fluorescence microscopy. Biocompatible molecular rulers are now available for the first time to calibrate modern super-resolution microscopy methods.
Recent advances in super-resolution microscopy have enabled optical resolution of several nanometers, comparable to the size of cellular molecules. However, it has been difficult to validate this resolution in cellular components such as multiprotein complexes, due to the lack of biomolecular reference systems capable of precise dye labeling at nanometer-scale distances.
Innovative biomolecular tools: PicoRulers
A team of scientists led by Dr. Gerti Bellew and Professor Markus Sauer from the Rudolf Wilhau Center for Integrative and Translational Bioimaging at the Julius Maximilian University (JMU) Würzburg in Bavaria, Germany, has now brought points.in a diary advanced materialsthey present a new biocompatible molecular ruler, PicoRulers (protein-based image calibration optical rulers).
The research team was able to construct these customized molecular rulers using genetic code expansion and click chemistry. These can be used as accurate biomolecular reference structures in fluorescence microscopy.
A masterpiece of technology: precision at the molecular scale
PicoRulers are based on the three-part protein PCNA (proliferating cell nuclear antigen). DNA Replication and repair. By accurately introducing unnaturalness, amino acid The protein is modified to allow fluorescent dyes or other molecules to click specifically at precisely defined positions and with minimal binding errors.
This will enable researchers to test the resolution of modern super-resolution microscopy techniques on precisely defined cellular biomolecules with unprecedented precision.
Markus Sauer enthuses: “The ability to resolve real biological structures at the sub-10 nanometer level heralds a new era in biological imaging. is characterized not only by its biological compatibility; it also enables unparalleled precision for testing resolution under realistic conditions.”
Broadening your research horizons: elucidating cellular processes
Applications of this technology go far beyond the limits of traditional microscopy. “Our PicoRulers are not only a tool for more precise measurements, but also open the door to deeper and more detailed investigation of the complex processes that occur within cells,” said Gerti Beliu. I will explain.
Implications for the future: Transforming biological imaging
Further development of PicoRulers has the potential to transform biological and medical imaging with molecular resolution in the long term. This makes it possible for the first time to validate and improve the resolution potential of new super-resolution microscopy methods for biological samples. This makes it a valuable tool for future elucidation of the molecular organization and interactions of biomolecules within cells.
Reference: “PCNAs as protein-based nanoruler for sub-10 Nm fluorescence imaging” by Dominic A. Helmerich, Made Budiarta, Danush Taban, Sören Doose, Gerti Beliu, Markus Sauer, November 27, 2023. advanced materials.