Smart light-sheet fluorescence microscopy

Background

The light-sheet fluorescence microscope is used to study three-dimensional samples combining the advantages of the speed of traditional wide-field epi-fluorescence microscopy with the optical sectioning power of a confocal microscope.  This combination makes it an ideal choice for the study of three-dimensional samples with high temporal dynamics.  The problem lies in that all biological samples are optical inhomogeneous leading to the introduction of phase aberrations that cause the images registered by these microscopes to be degraded and constitute a loss in information in the imaging process.  Through adaptive optical elements this missing information can be found and the true distribution of fluorescence can be elucidated.  

Objective

To find novel techniques that apply adaptive optics to light-sheet fluorescence microscopy in order to obtain better images and more information from biological samples.

Outcome

We have to date produced methodologies for changing the shape of the incident light to be better suited for light-sheet imaging; we have corrected aberrations in the illumination path and found a technique to correct for aberrations in the detected fluorescence light.

Project team members

  • Dean Wilding
  • Paolo Pozzi
  • Oleg Soloviev
  • Gleb Vdovin
  • Michel Verhaegen

Keywords

adaptive optics, microscopy, deconvolution

Sponsored by

European Research Council (ERC) – iCON Project

Partners

Erasmus MC

Universiteit Leiden

University of Texas Southwestern

Work programme

(1) Design and shaping of light for illumination.

(2) Correction of aberrations in illumination.

(3) Correction of aberrations in detection.

Selected publications

 

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