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PSF engineering in multifocus microscopy for increased depth volumetric imaging.

Bassam Hajj1, Mohamed El Beheiry2, Maxime Dahan3

  • 1Laboratoire Physico-Chimie, Institut Curie, PSL Research University, CNRS UMR168, 75005, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, 75005, Paris, France; Transcription Imaging Consortium, Janelia Research Campus, 19700 Helix Drive, Ashburn VA, 20147, USA; bassam.hajj@curie.fr.

Biomedical Optics Express
|May 28, 2016
PubMed
Summary
This summary is machine-generated.

We enhanced 3D single molecule imaging depth using multifocal microscopy and point spread function engineering. This breakthrough improves super-resolution imaging and molecule tracking in biological samples.

Keywords:
(100.6640) Superresolution(100.6890) Three-dimensional image processing(170.6900) Three-dimensional microscopy

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Accurate 3D imaging and localization of single molecules is crucial but technically demanding.
  • Existing methods face limitations in achieving sufficient depth for volumetric analysis.

Purpose of the Study:

  • To enhance the axial imaging depth for single molecule localization microscopy (SMLM).
  • To demonstrate the utility of the developed technique for super-resolution imaging and particle tracking in 3D.

Main Methods:

  • Integration of multifocal microscopy with engineered point spread functions (PSFs).
  • Utilized a novel approach to extend the depth range for volumetric single-molecule imaging.

Main Results:

  • Achieved 3D single molecule localization-based super-resolution imaging over an axial depth of 4 µm.
  • Successfully tracked diffusing nanoparticles in a fluid environment across an 8 µm depth range.

Conclusions:

  • The combined technique significantly extends the achievable depth for high-accuracy 3D single molecule imaging.
  • This advancement has broad implications for super-resolution microscopy and dynamic biological process studies in 3D.