title: Simple method for sub-diffraction resolution imaging of cellular structures on standard confocal microscopes by three-photon absorption of quantum dots.
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.
authors.affiliation: Confocal and 2-Photon Microscopy Core Facility, Max-Delbrueck-Center for Molecular Medicine Berlin, Berlin, Germany.
authors.forename: Anje
authors.initials: A
authors.lastname: Sporbert
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Cseresnyes
authors.forename: Meike
authors.initials: M
authors.lastname: Heidbreder
authors.forename: Petra
authors.initials: P
authors.lastname: Domaing
authors.forename: Stefan
authors.initials: S
authors.lastname: Hauser
authors.forename: Barbara
authors.initials: B
authors.lastname: Kaltschmidt
authors.forename: Christian
authors.initials: C
authors.lastname: Kaltschmidt
authors.forename: Mike
authors.initials: M
authors.lastname: Heilemann
authors.forename: Darius
authors.initials: D
authors.lastname: Widera
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007834:Lasers; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D017785:Photons; D045663:Quantum Dots