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Simple method for sub-diffraction resolution imaging of cellular structures on standard confocal microscopes by three-photon absorption of quantum dots

Authors

  • A. Sporbert
  • Z. Cseresnyes
  • M. Heidbreder
  • P. Domaing
  • S. Hauser
  • B. Kaltschmidt
  • C. Kaltschmidt
  • M. Heilemann
  • D. Widera

Journal

  • PLoS ONE

Citation

  • PLoS ONE 8 (5): e64023

Abstract

  • This study describes a simple technique that improves a recently developed 3D sub-diffraction imaging method based on three-photon absorption of commercially available quantum dots. The method combines imaging of biological samples via tri-exciton generation in quantum dots with deconvolution and spectral multiplexing, resulting in a novel approach for multi-color imaging of even thick biological samples at a 1.4 to 1.9-fold better spatial resolution. This approach is realized on a conventional confocal microscope equipped with standard continuous-wave lasers. We demonstrate the potential of multi-color tri-exciton imaging of quantum dots combined with deconvolution on viral vesicles in lentivirally transduced cells as well as intermediate filaments in three-dimensional clusters of mouse-derived neural stem cells (neurospheres) and dense microtubuli arrays in myotubes formed by stacks of differentiated C2C12 myoblasts.


DOI

doi:10.1371/journal.pone.0064023