Accelerated FRET-PAINT microscopy

Jongjin Lee; Sangjun Park; Sungchul Hohng

doi:10.1186/s13041-018-0414-3

Accelerated FRET-PAINT microscopy

Mol Brain. 2018 Nov 22;11(1):70. doi: 10.1186/s13041-018-0414-3.

Authors

Jongjin Lee¹, Sangjun Park¹, Sungchul Hohng^{2

3}

Affiliations

¹ Department of Physics and Astronomy, Seoul National University, Seoul, 08826, Republic of Korea.
² Department of Physics and Astronomy, Seoul National University, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.
³ Institute of Applied Physics, Seoul National University, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.

Abstract

Recent development of FRET-PAINT microscopy significantly improved the imaging speed of DNA-PAINT, the previously reported super-resolution fluorescence microscopy with no photobleaching problem. Here we try to achieve the ultimate speed limit of FRET-PAINT by optimizing the camera speed, dissociation rate of DNA probes, and bleed-through of the donor signal to the acceptor channel, and further increase the imaging speed of FRET-PAINT by 8-fold. Super-resolution imaging of COS-7 microtubules shows that high-quality 40-nm resolution images can be obtained in just tens of seconds.

Keywords: FRET; FRET-PAINT; Single-molecule localization microscopy; Super-resolution fluorescence microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
COS Cells
Chlorocebus aethiops
Fluorescence Resonance Energy Transfer*
Image Processing, Computer-Assisted
Microscopy, Fluorescence*
Nanotechnology
Signal-To-Noise Ratio