title: Axial tomography in live cell laser microscopy.
abstract: Single cell microscopy in a three-dimensional (3-D) environment is reported. Cells are grown in an agarose culture gel, located within microcapillaries and observed from different sides after adaptation of an innovative device for sample rotation. Thus, z -stacks can be recorded by confocal microscopy in different directions and used for illustration in 3-D. This gives additional information, since cells or organelles that appear superimposed in one direction, may be well resolved in another one. The method is tested and validated with single cells expressing a membrane or a mitochondrially associated green fluorescent protein, or cells accumulating fluorescent quantum dots. In addition, axial tomography supports measurements of cellular uptake and distribution of the anticancer drug doxorubicin in the nucleus (2 to 6 h after incubation) or the cytoplasm (24 h). This paper discusses that upon cell rotation an enhanced optical resolution in lateral direction compared to axial direction can be utilized to obtain an improved effective 3-D resolution, which represents an important step toward super-resolution microscopy of living cells.
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Richter
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Bruns
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bruns
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Weber
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Wagner
authors.affiliation: University of Heidelberg, Institute of Pharmacy and Molecular Biology, Im Neuenheimer Feld 364, 69120 Heidelberg, GermanycInstitute of Molecular Biology, Ackermannweg 4, 55128 Mainz, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Cremer
authors.affiliation: Aalen University, Institute of Applied Research, Beethovenstraße 1, 73430 Aalen, Germany.
authors.forename: Herbert
authors.initials: H
authors.lastname: Schneckenburger
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000295:instrumentation*; D004317:Doxorubicin / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal; D045663:Quantum Dots; D014057:Tomography, X-Ray Computed
chemical_list: D049452:Green Fluorescent Proteins; D004317:Doxorubicin