Optimizing imaging speed and excitation intensity for single-molecule localization microscopy

Nat Methods. 2020 Sep;17(9):909-912. doi: 10.1038/s41592-020-0918-5. Epub 2020 Aug 17.

Abstract

High laser powers are common practice in single-molecule localization microscopy to speed up data acquisition. Here we systematically quantified how excitation intensity influences localization precision and labeling density, the two main factors determining data quality. We found a strong trade-off between imaging speed and quality and present optimized imaging protocols for high-throughput, multicolor and three-dimensional single-molecule localization microscopy with greatly improved resolution and effective labeling efficiency.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbocyanines
  • Cell Line, Tumor
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Single Molecule Imaging / methods*
  • Time Factors

Substances

  • Alexa Fluor 647
  • Carbocyanines