Manuel F Juette

Publications (5)43.3 Total impact

• Article: Adaptive optics enables three-dimensional single particle tracking at the sub-millisecond scale

  Manuel F. Juette, Felix E. Rivera-Molina, Derek K. Toomre, Joerg Bewersdorf
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  ABSTRACT: We present the integration of an adaptive optics element into a feedback-driven single particle tracking microscope. Our instrument captures three-dimensional (3D) trajectories with down to 130 ls temporal resolution for dynamic studies on the nanoscale. Our 3D beam steering approach tracks particles over an axial range of >6 lm with �2ms mechanical response times and isolates the sample from any tracking motion. Tracking of transport vesicles containing Alexa488-labeled transferrin glycoprotein in living cells demonstrates the speed and sensitivity of our instrument. VC 2013 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 Unported License. [http://dx.doi.org/10.1063/1.4803538]
  Applied Physics Letters 04/2013; · 3.84 Impact Factor
• Article: Quantitative pupil analysis in stimulated emission depletion microscopy using phase retrieval.

  Emil B Kromann, Travis J Gould, Manuel F Juette, Jens E Wilhjelm, Joerg Bewersdorf
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  ABSTRACT: The resolution attainable with stimulated emission depletion (STED) microscopy greatly depends on the quality of the STED laser focus. So far, visual inspection of a measured STED focus has been the only convenient means of gauging the source of aberrations. Here we describe a method, requiring no instrument modifications, for obtaining an equivalent to the complex pupil function at the back aperture of the objective and show that it provides quantitative information about aberration sources (including aberrations induced by the objective or sample). We show the accuracy of this field representation to be sufficient for reconstructing the STED focus in three dimensions and determining corrective steps.
  Optics Letters 06/2012; 37(11):1805-7. · 3.40 Impact Factor
• Article: Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution.

  Manuel F Juette, Joerg Bewersdorf
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  ABSTRACT: Observing dynamics at the nanoscale requires submillisecond time resolution. Notably, in studying biological systems, three-dimensional (3D) trajectories of fluorescently labeled objects such as viruses or transport vesicles often need to be acquired with high temporal resolution. Here, we present a novel instrument that combines scanning-free multiplane detection at 3.2 kHz frame rate and single photon sensitivity with optimized beam-steering capabilities. This setup enables ultrafast 3D localization with submillisecond time resolution and nanometer localization precision. We demonstrate 3D tracking of single fluorescent particles at speeds of up to 150 nm/ms over several seconds and large volumes. By focused excitation of only the particle of interest, while avoiding confocal pinholes, the system realizes maximum detection efficiency at minimal laser irradiation. These features, combined with the avoidance of stage movement, provide high live-sample compatibility for future biomedical applications.
  Nano Letters 10/2010; 10(11):4657-63. · 13.20 Impact Factor
• Article: Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy.

  Michael J Mlodzianoski, Manuel F Juette, Glen L Beane, Joerg Bewersdorf
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  ABSTRACT: Three-dimensional (3D) particle localization at the nanometer scale plays a central role in 3D particle tracking and 3D localization-based super-resolution microscopy. Here we introduce a localization algorithm that is independent of theoretical models and therefore generally applicable to a large number of experimental realizations. Applying this algorithm and a convertible experimental setup we compare the performance of the two major 3D techniques based on astigmatic distortions and on multiplane detection. In both methods we obtain experimental 3D localization accuracies in agreement with theoretical predictions and characterize the depth dependence of the localization accuracy in detail.
  Optics Express 06/2009; 17(10):8264-77. · 3.59 Impact Factor
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  Article: Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples.

  Manuel F Juette, Travis J Gould, Mark D Lessard, Michael J Mlodzianoski, Bhupendra S Nagpure, Brian T Bennett, Samuel T Hess, Joerg Bewersdorf
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  ABSTRACT: Imaging volumes as thick as whole cells at three-dimensional (3D) super-resolution is required to reveal unknown features of cellular organization. We report a light microscope that generates images with translationally invariant 30 x 30 x 75 nm resolution over a depth of several micrometers. This method, named biplane (BP) FPALM, combines a double-plane detection scheme with fluorescence photoactivation localization microscopy (FPALM) enabling 3D sub-diffraction resolution without compromising speed or sensitivity.
  Nature Methods 07/2008; 5(6):527-9. · 19.28 Impact Factor