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Fluorescent molecules emit light in a dipole radiation pattern that can be used to infer their orientation through defocused fluorescence microscopy. Proper measurement of the orientation requires mathematical modeling of the radiation…

Optics · Physics 2025-08-07 E. Dey , M. Elorza , F. W. Foss , J. J. Gomez Cadenas , B. J. P. Jones

We demonstrate theoretically and experimentally that the three-dimensional orientation of a single fluorescent nano-emitter can be determined by polarization analysis of the emitted light (while excitation polarization analysis provides…

Fluorescent molecules are versatile nanoscale emitters that enable detailed observations of biophysical processes with nanoscale resolution. Because they are well-approximated as electric dipoles, imaging systems can be designed to…

Optics · Physics 2024-03-26 Tingting Wu , Matthew D. Lew

Optical fluorescence imaging is capable of measuring both the spatial and rotational dynamics of single molecules. However, unavoidable measurement noise will result in inaccurate estimates of rotational dynamics, causing a molecule to…

Optics · Physics 2019-05-22 Oumeng Zhang , Matthew D. Lew

Precisely measuring the three-dimensional position and orientation of individual fluorophores is challenging due to the substantial photon shot noise in single-molecule experiments. Facing this limited photon budget, numerous techniques…

Optics · Physics 2021-02-02 Oumeng Zhang , Matthew D. Lew

When circularly polarized light is scattered from a rotating target, a rotational Doppler shift (RDS) emerges from an exchange of angular momentum between the spinning object and the electromagnetic field. Here, we used coherently spinning…

We present high resolution two-photon excitation microscopy studies combining two-photon fluorescence (TPF) and second harmonic generation (SHG) in order to probe orientational distributions of molecular ensembles at room temperature. A…

Soft Condensed Matter · Physics 2007-05-23 Veronique Le Floc'h , Sophie Brasselet , Jean-Francois Roch , Joseph Zyss

We have developed and experimentally demonstrated an improved electrostatic deflector for the spatial separation of molecules according to their dipole-moment-to-mass ratio. The device features a very open structure that allows for…

Chemical Physics · Physics 2017-07-25 Jens S. Kienitz , Karol Długołecki , Sebastian Trippel , Jochen Küpper

Single molecule imaging is one of the main target areas of X-ray free electron lasers. It relies on the possibility of orienting the large number of low counting statistics 2D diffraction patterns taken at random orientations of identical…

Atomic and Molecular Clusters · Physics 2012-11-30 Zoltan Jurek , Gyula Faigel

We investigate rotational diffusion of fluorescent molecules in angular potential wells, the excitation and subsequent emissions from these diffusing molecules, and the imaging of these emissions with high-NA aplanatic optical microscopes.…

Measuring subdiffraction separations between single fluorescent particles is important for biological, nano-, and medical-technology studies. Major challenges include (i) measuring changing molecular separations with high temporal…

Biological Physics · Physics 2015-05-18 Shawn H. DeCenzo , Michael C. DeSantis , Y. M. Wang

In single-molecule super-resolution microscopy, engineered point-spread functions (PSFs) are designed to efficiently encode new molecular properties, such as 3D orientation, into complex spatial features captured by a camera. To fully…

Image and Video Processing · Electrical Eng. & Systems 2019-07-16 Hesam Mazidi , Eshan S. King , Oumeng Zhang , Arye Nehorai , Matthew D. Lew

We consider deflection of rotating symmetric molecules by inhomogeneous optical and static electric fields, compare results with the case of linear molecules, and find new singularities in the distribution of the scattering angle.…

Chemical Physics · Physics 2015-05-28 Erez Gershnabel , Ilya Sh. Averbukh

Various techniques have been developed to measure the 2D and 3D positions and 2D and 3D orientations of fluorescent molecules with improved precision over standard epifluorescence microscopes. Due to the challenging signal-to-background…

Optics · Physics 2021-02-02 Oumeng Zhang , Matthew D. Lew

Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging.…

We provide a theory of the deflection of polar and non-polar rotating molecules by inhomogeneous static electric field. Rainbow-like features in the angular distribution of the scattered molecules are analyzed in detail. Furthermore, we…

Chemical Physics · Physics 2015-05-20 E. Gershnabel , I. Sh. Averbukh

Single fluorescent molecules, behaving as ideal electric dipole emitters, are powerful nanoscopic probes of complex optical fields. Here, this property is exploited to precisely map the polarization and vectorial structure of tightly…

A beam of rotating dipolar particles (molecules or clusters) will broaden when passed through an electric or magnetic field gradient region. This broadening, which is a common experimental observable, can be expressed in terms of the…

Atomic and Molecular Clusters · Physics 2012-01-05 Jaap Bulthuis , Vitaly V. Kresin

The mixed-field orientation of an asymmetric-rotor molecule with its permanent dipole moment non-parallel to the principal axes of polarizability is investigated experimentally and theoretically. We find that for the typical case of a…

The rich information content of measurements in the molecular frame rather than the laboratory frame has motivated the development of several methods for aligning gas phase molecules in space. Even so, for asymmetric tops the problem of…

Atomic Physics · Physics 2016-11-23 Varun Makhija , Xiaoming Ren , Drue Gockel , Anh-Thu Le , Vinod Kumarappan
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