Single-fluorophore Orientation Determination with Multiview Polarized Illumination: Modeling and Microscope Design

Overview

Journal Opt Express

Specialties Biophysics
Ophthalmology

Date 2017 Dec 17

PMID 29245807

Citations 5

Authors

Talon Chandler

Shalin Mehta

Hari Shroff

Rudolf Oldenbourg

Patrick J La Riviere

Affiliations

Soon will be listed here.

Abstract

We investigate the use of polarized illumination in multiview microscopes for determining the orientation of single-molecule fluorescence transition dipoles. First, we relate the orientation of single dipoles to measurable intensities in multiview microscopes and develop an information-theoretic metric-the solid-angle uncertainty-to compare the ability of multiview microscopes to estimate the orientation of single dipoles. Next, we compare a broad class of microscopes using this metric-single- and dual-view microscopes with varying illumination polarization, illumination numerical aperture (NA), detection NA, obliquity, asymmetry, and exposure. We find that multi-view microscopes can measure all dipole orientations, while the orientations measurable with single-view microscopes is halved because of symmetries in the detection process. We also find that choosing a small illumination NA and a large detection NA are good design choices, that multiview microscopes can benefit from oblique illumination and detection, and that asymmetric NA microscopes can benefit from exposure asymmetry.

Citing Articles

Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope.

Chandler T, Guo M, Su Y, Chen J, Wu Y, Liu J Proc Natl Acad Sci U S A. 2025; 122(8):e2406679122.

PMID: 39982748 PMC: 11874040. DOI: 10.1073/pnas.2406679122.

Permittivity tensor imaging: modular label-free imaging of 3D dry mass and 3D orientation at high resolution.

Yeh L, Ivanov I, Chandler T, Byrum J, Chhun B, Guo S Nat Methods. 2024; 21(7):1257-1274.

PMID: 38890427 PMC: 11239526. DOI: 10.1038/s41592-024-02291-w.

Three-dimensional spatio-angular fluorescence microscopy with a polarized dual-view inverted selective-plane illumination microscope (pol-diSPIM).

Chandler T, Guo M, Su Y, Chen J, Wu Y, Liu J bioRxiv. 2024; .

PMID: 38712306 PMC: 11071302. DOI: 10.1101/2024.03.09.584243.

Photon efficient orientation estimation using polarization modulation in single-molecule localization microscopy.

Thorsen R, Hulleman C, Rieger B, Stallinga S Biomed Opt Express. 2022; 13(5):2835-2858.

PMID: 35774337 PMC: 9203119. DOI: 10.1364/BOE.452159.

Dipole-spread-function engineering for simultaneously measuring the 3D orientations and 3D positions of fluorescent molecules.

Wu T, Lu J, Lew M Optica. 2022; 9(5):505-511.

PMID: 35601691 PMC: 9122034. DOI: 10.1364/OPTICA.451899.

References

Mehta S, McQuilken M, La Riviere P, Occhipinti P, Verma A, Oldenbourg R . Dissection of molecular assembly dynamics by tracking orientation and position of single molecules in live cells. Proc Natl Acad Sci U S A. 2016; 113(42):E6352-E6361. PMC: 5081662. DOI: 10.1073/pnas.1607674113. View

Fourkas J . Rapid determination of the three-dimensional orientation of single molecules. Opt Lett. 2007; 26(4):211-3. DOI: 10.1364/ol.26.000211. View

Wu Y, Chandris P, Winter P, Kim E, Jaumouille V, Kumar A . Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy. Optica. 2016; 3(8):897-910. PMC: 5066810. DOI: 10.1364/OPTICA.3.000897. View

Wu Y, Kumar A, Smith C, Ardiel E, Chandris P, Christensen R . Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy. Nat Commun. 2017; 8(1):1452. PMC: 5682293. DOI: 10.1038/s41467-017-01250-8. View

Toprak E, Enderlein J, Syed S, McKinney S, Petschek R, Ha T . Defocused orientation and position imaging (DOPI) of myosin V. Proc Natl Acad Sci U S A. 2006; 103(17):6495-9. PMC: 1458912. DOI: 10.1073/pnas.0507134103. View

Peterman E, Sosa H, Goldstein L, Moerner W . Polarized fluorescence microscopy of individual and many kinesin motors bound to axonemal microtubules. Biophys J. 2001; 81(5):2851-63. PMC: 1301750. DOI: 10.1016/S0006-3495(01)75926-7. View

Backer A, Moerner W . Extending single-molecule microscopy using optical Fourier processing. J Phys Chem B. 2014; 118(28):8313-29. PMC: 4317050. DOI: 10.1021/jp501778z. View

Lu C, Vanden Bout D . Analysis of orientational dynamics of single fluorophore trajectories from three-angle polarization experiments. J Chem Phys. 2008; 128(24):244501. DOI: 10.1063/1.2937730. View

Chhetri R, Amat F, Wan Y, Hockendorf B, Lemon W, Keller P . Whole-animal functional and developmental imaging with isotropic spatial resolution. Nat Methods. 2015; 12(12):1171-8. DOI: 10.1038/nmeth.3632. View

10.

Backlund M, Lew M, Backer A, Sahl S, Moerner W . The role of molecular dipole orientation in single-molecule fluorescence microscopy and implications for super-resolution imaging. Chemphyschem. 2014; 15(4):587-99. PMC: 3992256. DOI: 10.1002/cphc.201300880. View

11.

Anantharam A, Onoa B, Edwards R, Holz R, Axelrod D . Localized topological changes of the plasma membrane upon exocytosis visualized by polarized TIRFM. J Cell Biol. 2010; 188(3):415-28. PMC: 2819686. DOI: 10.1083/jcb.200908010. View

12.

McQuilken M, Jentzsch M, Verma A, Mehta S, Oldenbourg R, Gladfelter A . Analysis of Septin Reorganization at Cytokinesis Using Polarized Fluorescence Microscopy. Front Cell Dev Biol. 2017; 5:42. PMC: 5413497. DOI: 10.3389/fcell.2017.00042. View

13.

Agrawal A, Quirin S, Grover G, Piestun R . Limits of 3D dipole localization and orientation estimation for single-molecule imaging: towards Green's tensor engineering. Opt Express. 2012; 20(24):26667-80. DOI: 10.1364/OE.20.026667. View

14.

Wu Y, Wawrzusin P, Senseney J, Fischer R, Christensen R, Santella A . Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy. Nat Biotechnol. 2013; 31(11):1032-8. PMC: 4105320. DOI: 10.1038/nbt.2713. View

15.

DeMay B, Noda N, Gladfelter A, Oldenbourg R . Rapid and quantitative imaging of excitation polarized fluorescence reveals ordered septin dynamics in live yeast. Biophys J. 2011; 101(4):985-94. PMC: 3175061. DOI: 10.1016/j.bpj.2011.07.008. View

16.

Shribak M, Oldenbourg R . Techniques for fast and sensitive measurements of two-dimensional birefringence distributions. Appl Opt. 2003; 42(16):3009-17. DOI: 10.1364/ao.42.003009. View

17.

Tyo J, Goldstein D, Chenault D, Shaw J . Review of passive imaging polarimetry for remote sensing applications. Appl Opt. 2006; 45(22):5453-69. DOI: 10.1364/ao.45.005453. View

18.

Backer A, Lee M, Moerner W . Enhanced DNA imaging using super-resolution microscopy and simultaneous single-molecule orientation measurements. Optica. 2016; 3(6):3-6. PMC: 5050005. DOI: 10.1364/OPTICA.3.000659. View

19.

Forkey J, Quinlan M, Shaw M, Corrie J, Goldman Y . Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization. Nature. 2003; 422(6930):399-404. DOI: 10.1038/nature01529. View