title: Single molecule microscopy in 3D cell cultures and tissues.
abstract: From the onset of the first microscopic visualization of single fluorescent molecules in living cells at the beginning of this century, to the present, almost routine application of single molecule microscopy, the method has well-proven its ability to contribute unmatched detailed insight into the heterogeneous and dynamic molecular world life is composed of. Except for investigations on bacteria and yeast, almost the entire story of success is based on studies on adherent mammalian 2D cell cultures. However, despite this continuous progress, the technique was not able to keep pace with the move of the cell biology community to adapt 3D cell culture models for basic research, regenerative medicine, or drug development and screening. In this review, we will summarize the progress, which only recently allowed for the application of single molecule microscopy to 3D cell systems and give an overview of the technical advances that led to it. While initially posing a challenge, we finally conclude that relevant 3D cell models will become an integral part of the on-going success of single molecule microscopy.
authors.affiliation: Membrane Dynamics, Department of Biology, Technische Universität Darmstadt, Schnittspahnstrasse 3-5, 64287 Darmstadt, Germany.
authors.forename: Florian M
authors.initials: FM
authors.lastname: Lauer
authors.affiliation: Membrane Dynamics, Department of Biology, Technische Universität Darmstadt, Schnittspahnstrasse 3-5, 64287 Darmstadt, Germany; Institute of Health and Biomedical Innovation, Science and Engineering Faculty, Queensland University of Technology, 60 Musk Ave, Kelvin Grove, 4059 QLD, Brisbane, Australia.
authors.forename: Elke
authors.initials: E
authors.lastname: Kaemmerer
authors.affiliation: Membrane Dynamics, Department of Biology, Technische Universität Darmstadt, Schnittspahnstrasse 3-5, 64287 Darmstadt, Germany. Electronic address: tobias.meckel@me.com.
authors.forename: Tobias
authors.initials: T
authors.lastname: Meckel
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D015195:Drug Design; D006801:Humans; D008853:Microscopy / Q000379:methods*; D008954:Models, Biological; D044968:Regenerative Medicine / Q000379:methods
keywords: 3D cell cultures; Adaptive optics; HILO illumination; Light sheet illumination; Multiphoton microscopy; Single molecule microscopy; Spheroids; Two-photon microscopy