Adaptive optics in single objective inclined light sheet microscopy enables three-dimensional localization microscopy in adult Drosophila brains
Shih Te Hung (TU Delft - Team Carlas Smith)
Arnau Llobet Rosell (University of Lausanne)
Daphne Jurriens (Universiteit Utrecht)
O.A. Soloviev (TU Delft - Team Michel Verhaegen)
Lukas C. Kapitein (Universiteit Utrecht)
Kristin S. Grußmayer (Kavli institute of nanoscience Delft, TU Delft - BN/Kristin Grussmayer Lab)
Lukas J. Neukomm (University of Lausanne)
M. Verhaegen (TU Delft - Team Michel Verhaegen)
C.S. Smith (TU Delft - Team Carlas Smith, ImPhys/Computational Imaging)
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Abstract
Single-molecule localization microscopy (SMLM) enables the high-resolution visualization of organelle structures and the precise localization of individual proteins. However, the expected resolution is not achieved in tissue as the imaging conditions deteriorate. Sample-induced aberrations distort the point spread function (PSF), and high background fluorescence decreases the localization precision. Here, we synergistically combine sensorless adaptive optics (AO), in-situ 3D-PSF calibration, and a single-objective lens inclined light sheet microscope (SOLEIL), termed (AO-SOLEIL), to mitigate deep tissue-induced deteriorations. We apply AO-SOLEIL on several dSTORM samples including brains of adult Drosophila. We observed a 2x improvement in the estimated axial localization precision with respect to widefield without aberration correction while we used synergistic solution. AO-SOLEIL enhances the overall imaging resolution and further facilitates the visualization of sub-cellular structures in tissue.
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