Optical Sectioning and High Resolution in Single-Slice Structured Illumination Microscopy by Thick Slice Blind-SIM Reconstruction

Aurélie Jost (1, 2) , Elen Tolstik (1, 2) , Polina Feldmann (1, 2) , Kai Wicker (1, 2, 3) , Anne Sentenac (4) , Rainer Heintzmann (5, 1, 2)

1 Institute of Physical Chemistry, Abbe Center of Photonics
2 Leibniz IPHT - Leibniz Institute of Photonic Technology
3 Carl Zeiss AG
4 SEMO - SEMO
5 King‘s College London

The microscope image of a thick fluorescent sample taken at a given focal plane is plagued by out-of-focus fluorescence and diffraction limited resolution. In this work, we show that a single slice of Structured Illumination Microscopy (two or three beam SIM) data can be processed to provide an image exhibiting tight sectioning and high transverse resolution. Our reconstruction algorithm is adapted from the blind-SIM technique which requires very little knowledge of the illumination patterns. It is thus able to deal with illumination distortions induced by the sample or illumination optics. We named this new algorithm thick slice blind-SIM because it models a three-dimensional sample even though only a single two-dimensional plane of focus was measured.

Mots clés: FLUORESCENCE MICROSCOPY; LIGHT

Domaines: Sciences du Vivant [q-bio]; Imagerie; Optique [physics.optics]; Optique / photonique; Traitement du signal et de l'image [eess.SP]

Aurélie Jost, Elen Tolstik, Polina Feldmann, Kai Wicker, Anne Sentenac, et al.. Optical Sectioning and High Resolution in Single-Slice Structured Illumination Microscopy by Thick Slice Blind-SIM Reconstruction. PLoS ONE, 2015, 10, pp.e0132174. ⟨10.1371/journal.pone.0132174⟩. ⟨hal-01199875⟩