Quantification of the NA dependent change of shape in the image formation of a z-polarized fluorescent molecule using vectorial diffraction simulations.
Authors
Bruggeman, Ezra
Rowlands, Christopher J
Wolfson, Deanna L
Ahluwalia, Balpreet S
Publication Date
2022-05Journal Title
Microsc Res Tech
ISSN
1059-910X
Publisher
Wiley
Volume
85
Issue
5
Pages
2016-2022
Language
en
Type
Article
This Version
AO
VoR
Metadata
Show full item recordCitation
Ströhl, F., Bruggeman, E., Rowlands, C. J., Wolfson, D. L., & Ahluwalia, B. S. (2022). Quantification of the NA dependent change of shape in the image formation of a z-polarized fluorescent molecule using vectorial diffraction simulations.. Microsc Res Tech, 85 (5), 2016-2022. https://doi.org/10.1002/jemt.24060
Description
Funder: FP7 People: Marie‐Curie Actions; Id: http://dx.doi.org/10.13039/100011264; Grant(s): 836355
Funder: UiT Open Access fund
Funder: UiT Aurora Outstanding program
Abstract
The point spread function of a fixed fluorophore with its dipole axis colinear to the optical axis appears donut-shaped when seen through a microscope, and its light distribution in the pupil plane is radially polarized. Yet other techniques, such as photolithography, report that this same light distribution in the pupil plane appears as a solid spot. How can this same distribution lead to a spot in one case but a donut in the other? Here, we show how the tube lens of the system plays a critical role in determining this shape. Using a vectorial treatment of image formation, we simulate the relative contributions of both longitudinal and radial components to the image of a dipole emitter and thus show how the donut (typically reported for z-polarized single molecule fluorescence microscopy) transforms into a solid spot (as commonly reported for photolithography) as the numerical aperture of the tube lens increases. We find that the transition point occurs around 0.7 NA, which is significantly higher than used for most microscopy systems and lower than for common photolithography systems, thus resolving the seeming paradox of dipole shape.
Keywords
PRIMERS IN MICROSCOPY, image formation theory, Jones matrix calculus, microscopy, vectorial diffraction
Sponsorship
Norges Forskningsråd (314546)
Identifiers
jemt24060
External DOI: https://doi.org/10.1002/jemt.24060
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336542
Rights
Licence:
http://creativecommons.org/licenses/by/4.0/
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