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  4Pi MINFLUX arrangement maximizes spatio-temporal localization precision of fluorescence emitter

Rickert, J. D., Held, M. O., Engelhardt, J., & Hell, S. W. (2024). 4Pi MINFLUX arrangement maximizes spatio-temporal localization precision of fluorescence emitter. PNAS, 121(11): e2318870121, pp. 1-7. doi:10.1073/pnas.2318870121.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000E-A19B-D Version Permalink: https://hdl.handle.net/21.11116/0000-000E-A19C-C
Genre: Journal Article

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 Creators:
Rickert, Julian D.1, 2, Author           
Held, Marcus O.1, Author           
Engelhardt, Johann1, Author           
Hell, Stefan W.1, Author                 
Affiliations:
1Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society, ou_2364730              
2Max Planck School Matter to Life, Max Planck Schools, Max Planck Society, ou_3473638              

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Free keywords: MINFLUX localization; diffusion; fluorescence nanoscopy; superresolution microscopy
 Abstract: We introduce MINFLUX localization with interferometric illumination through opposing objective lenses for maximizing the attainable precision in 3D-localization of single inelastic scatterers, such as fluorophores. Our 4Pi optical configuration employs three sequentially tilted counter-propagating beam pairs for illumination, each providing a narrow interference minimum of illumination intensity at the focal point. The localization precision is additionally improved by adding the inelastically scattered or fluorescence photons collected through both objective lenses. Our 4Pi configuration yields the currently highest precision per detected photon among all localization schemes. Tracking gold nanoparticles as non-blinking inelastic scatterers rendered a position uncertainty <0.4 nm3 in volume at a localization frequency of 2.9 kHz. We harnessed the record spatio-temporal precision of our 4Pi MINFLUX approach to examine the diffusion of single fluorophores and fluorescent nanobeads in solutions of sucrose in water, revealing local heterogeneities at the nanoscale. Our results show the applicability of 4Pi MINFLUX to study molecular nano-environments of diffusion and its potential for quantifying rapid movements of molecules in cells and other material composites.

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Language(s): eng - English
 Dates: 2023-10-312024-01-302024-03-05
 Publication Status: Published online
 Pages: 7
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1073/pnas.2318870121
URI: https://pubmed.ncbi.nlm.nih.gov/38442172/
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Title: PNAS
  Other : Proceedings of the National Academy of Sciences of the United States of America
  Other : Proceedings of the National Academy of Sciences of the USA
  Abbreviation : Proc. Natl. Acad. Sci. U. S. A.
Source Genre: Journal
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Publ. Info: Washington, D.C. : National Academy of Sciences
Pages: - Volume / Issue: 121 (11) Sequence Number: e2318870121 Start / End Page: 1 - 7 Identifier: ISSN: 0027-8424
CoNE: https://pure.mpg.de/cone/journals/resource/954925427230