title: Preparation strategy and illumination of three-dimensional cell cultures in light sheet-based fluorescence microscopy.
abstract: A device for selective plane illumination microscopy (SPIM) of three-dimensional multicellular spheroids, in culture medium under stationary or microfluidic conditions, is described. Cell spheroids are located in a micro-capillary and a light sheet, for illumination, is generated in an optical setup adapted to a conventional inverse microscope. Layers of the sample, of about 10 μm or less in diameter, are, thus, illuminated selectively and imaged by high resolution fluorescence microscopy. SPIM is operated at low light exposure even if a larger number of layers is imaged and is easily combined with laser scanning microscopy. Chinese hamster ovary cells expressing a membrane-associated green fluorescent protein are used for preliminary tests, and the uptake of the fluorescent marker, acridine orange via a microfluidic system, is visualized to demonstrate its potential in cancer research such as for the detection of cellular responses to anticancer drugs.
authors.affiliation: Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, 73430 Aalen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bruns
authors.forename: Sarah
authors.initials: S
authors.lastname: Schickinger
authors.forename: Rainer
authors.initials: R
authors.lastname: Wittig
authors.forename: Herbert
authors.initials: H
authors.lastname: Schneckenburger
mesh_terms: D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry; D008027:Light; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D008856:Microscopy, Fluorescence / Q000379:methods*; D018874:Spheroids, Cellular / Q000737:chemistry* / Q000166:cytology*
chemical_list: D005456:Fluorescent Dyes