title: High-resolution deep imaging of live cellular spheroids with light-sheet-based fluorescence microscopy.
abstract: Conventional two-dimensional cell monolayers do not provide the geometrical, biochemical and mechanical cues found in real tissues. Cells in real tissues interact through chemical and mechanical stimuli with adjacent cells and via the extracellular matrix. Such a highly interconnected communication network extends along all three dimensions. This architecture is lost in two-dimensional cultures. Therefore, at least in many cases, two-dimensional cell monolayers do not represent a suitable in vitro tool to characterize accurately the biology of real tissues. Many studies performed over the last few years have demonstrated that the differences between three-dimensional and two-dimensional cultured cells are striking at the morphological and molecular levels and that three-dimensional cell cultures can be employed in order to shrink the gap between real tissues and in vitro cell models. End-point and long-term imaging of cellular and sub-cellular processes with fluorescence microscopy provides direct insight into the physiological behavior of three-dimensional cell cultures and their response to chemical or mechanical stimulation. Fluorescence imaging of three-dimensional cell cultures sets new challenges and imposes specific requirements concerning the choice of a suitable microscopy technique. Deep penetration into the specimen, high imaging speed and ultra-low intensity of the excitation light are key requirements. Light-sheet-based fluorescence microscopy (LSFM) offers a favorable combination of these requirements and is therefore currently established as the technique of choice for the study of three-dimensional cell cultures. This review illustrates the benefits of cellular spheroids in the life sciences and suggests that LSFM is essential for investigations of cellular and sub-cellular dynamic processes in three-dimensions over time and space.
authors: Buchmann Institute for Molecular Life Sciences (BMLS), Goethe Universität Frankfurt am Main, Max-von-Laue-Straße 15, 60438, Frankfurt am Main, Germany.
authors: Francesco
authors: F
authors: Pampaloni
authors: Nariman
authors: N
authors: Ansari
authors: Ernst H K
authors: EH
authors: Stelzer
mesh_terms: D000818:Animals; D002450:Cell Communication; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D004867:Equipment Design; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D018874:Spheroids, Cellular / Q000166:cytology* / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure*