Single-molecule orientation localization microscopy II: a performance comparison

Oumeng Zhang, Matthew D. Lew

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background ratio in typical single-molecule experiments, it is essential to choose an imaging system optimized for the specific target sample. In this work, we compare the performance of multiple state-of-the-art and commonly used methods for orientation localization microscopy against the fundamental limits of measurement precision. Our analysis reveals optimal imaging methods for various experiment conditions and sample geometries. Interestingly, simple modifications to the standard fluorescence microscope exhibit superior performance in many imaging scenarios.

Original languageEnglish
Pages (from-to)288-297
Number of pages10
JournalJournal of the Optical Society of America A: Optics and Image Science, and Vision
Issue number2
StatePublished - Feb 2021


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