Three-Dimensional Single-Molecule Localization Microscopy in Whole-Cell and Tissue Specimens

Sheng Liu, Hyun Huh, Sang Hyuk Lee, Fang Huang

Research output: Contribution to journalReview articlepeer-review

16 Scopus citations


Super-resolution microscopy techniques are versatile and powerful tools for visualizing organelle structures, interactions, and protein functions in biomedical research. However, whole-cell and tissue specimens challenge the achievable resolution and depth of nanoscopy methods. We focus on three-dimensional single-molecule localization microscopy and review some of the major roadblocks and developing solutions to resolving thick volumes of cells and tissues at the nanoscale in three dimensions. These challenges include background fluorescence, system-and sample-induced aberrations, and information carried by photons, as well as drift correction, volume reconstruction, and photobleaching mitigation. We also highlight examples of innovations that have demonstrated significant breakthroughs in addressing the abovementioned challenges together with their core concepts as well as their trade-offs.

Original languageEnglish (US)
Pages (from-to)155-184
Number of pages30
JournalAnnual Review of Biomedical Engineering
StatePublished - Jun 4 2020

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • Biomedical Engineering


  • CRLB
  • Cramér-Rao lower bound
  • Super-resolution microscopy
  • adaptive optics
  • fluorescence microscopy
  • light-sheet microscopy
  • tissue imaging


Dive into the research topics of 'Three-Dimensional Single-Molecule Localization Microscopy in Whole-Cell and Tissue Specimens'. Together they form a unique fingerprint.

Cite this