Impact of optical aberrations on axial position determination by photometry
RNA Therapeutics Institute
Biochemistry, Biophysics, and Structural Biology | Bioimaging and Biomedical Optics
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
Nat Methods. 2018 Dec;15(12):989-990. doi: 10.1038/s41592-018-0227-4. Link to article on publisher's site
Thorsen, Rasmus O.; Hulleman, Christiaan N.; Hammer, Mathias; Grünwald, David; Stallinga, Sjoerd; and Rieger, Bernd, "Impact of optical aberrations on axial position determination by photometry" (2018). RNA Therapeutics Institute Publications. 58.