RNA Therapeutics Institute Publications

Title

Impact of optical aberrations on axial position determination by photometry

UMMS Affiliation

RNA Therapeutics Institute

Publication Date

2018-12-01

Document Type

Article

Disciplines

Biochemistry, Biophysics, and Structural Biology | Bioimaging and Biomedical Optics

Abstract

Recently, Franke et al.introduced a way to estimate the axial position of single molecules (temporal radial-aperture-based intensity estimation (TRABI)). To this end, they compared the detected photon count from a TRABI estimation to the estimated count from Gaussian point-spread function (PSF) fitting to the data. Empirically they found that the photometric ratio is ~0.7–0.8 at points close to focus and decreases as the distance from the focal plane increases. Here we explain this reported but unexplained discrepancy and, furthermore, show that the photometric ratio as an indicator for axial position is susceptible even to typical optical aberrations.

DOI of Published Version

10.1038/s41592-018-0227-4

Source

Nat Methods. 2018 Dec;15(12):989-990. doi: 10.1038/s41592-018-0227-4. Link to article on publisher's site

Journal/Book/Conference Title

Nature methods

Related Resources

Link to Article in PubMed

PubMed ID

30504865

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