title: Super-resolved 3-D imaging of live cells' organelles from bright-field photon transmission micrographs.
abstract: Current biological and medical research is aimed at obtaining a detailed spatiotemporal map of a live cell's interior to describe and predict cell's physiological state. We present here an algorithm for complete 3-D modelling of cellular structures from a z-stack of images obtained using label-free wide-field bright-field light-transmitted microscopy. The method visualizes 3-D objects with a volume equivalent to the area of a camera pixel multiplied by the z-height. The computation is based on finding pixels of unchanged intensities between two consecutive images of an object spread function. These pixels represent strongly light-diffracting, light-absorbing, or light-emitting objects. To accomplish this, variables derived from Rényi entropy are used to suppress camera noise. Using this algorithm, the detection limit of objects is only limited by the technical specifications of the microscope setup-we achieve the detection of objects of the size of one camera pixel. This method allows us to obtain 3-D reconstructions of cells from bright-field microscopy images that are comparable in quality to those from electron microscopy images.
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic. Electronic address: rrychtarikova@frov.jcu.cz.
authors.forename: Renata
authors.initials: R
authors.lastname: Rychtáriková
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Náhlík
authors.affiliation: Princeton University, Princeton, New Jersey 08544, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Shi
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
authors.forename: Daria
authors.initials: D
authors.lastname: Malakhova
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Macháček
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California 94305, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Smaha
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Urban
authors.affiliation: Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
authors.forename: Dalibor
authors.initials: D
authors.lastname: Štys
mesh_terms: D000465:Algorithms; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D015388:Organelles / Q000502:physiology*
keywords: 3D; Bright-field microscopy; Light transmission; Live cell imaging; Super-resolution