第1236回生物科学セミナー
Tracing chromosomes at the nanoscale with DNA-PAINT
Dr. Hiroshi Sasaki（Wyss Institute, Harvard University）
2018年09月06日(木) 16:00-17:00 理学部1号館東棟 380号室
Super-resolution microscopy has the potential to transform our understanding of the organization of the genome in individual cells. Localization microscopy techniques such as STORM [1] and DNA-PAINT [2] can resolve structures on the order of tens of nanometers size, providing rich structural detail about the organization and structure of biomolecules in fixed samples. We have harnessed highly multiplexed DNA-PAINT [3] to image chromosomes labeled by the Oligopaints FISH technique [4, 5]. We have optimized DNA-PAINT to work with a simple and easy to implement optical setup that includes an adaptive optics unit, and with this, we have achieved >10-color super-resolution images of human chromosomes at <15 nm lateral (X-Y) resolution and <25 nm axial (Z). We are now applying our imaging platform to visualize the structure of sub-chromosomal domains in human fibroblast cells and working towards >20-color imaging. Our approach will enable researchers to uncover the trajectory and nanoscale substructure of interphase chromosomes in single cells.
参考文献
1. Rust, Bates, and Zhuang. 2006. “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).” Nat. Methods 3(10):793–796.
2. Jungmann, et al., 2010. “Single-molecule kinetics and super-resolution microscopy by fluorescence imaging of transient binding on DNA origami.” Nano Lett. 10(11):4756–61.
3. Jungmann, et al., 2014. “Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT.” Nat. Methods 11(3):313–8.
4. Beliveau, et al., 2012. “Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes.” Proc. Natl. Acad. Sci. U S A. 109(52):21301–6.
5. Beliveau, et al., 2018. “OligoMiner provides a rapid, flexible environment for the design of genome-scale oligonucleotide in situ hybridization probes.” Proc. Natl. Acad. Sci. U S A. 115(10):E2183–E2192.
