Format

Send to

Choose Destination
Ultramicroscopy. 2017 Aug;179:1-14. doi: 10.1016/j.ultramic.2017.03.018. Epub 2017 Mar 18.

Super-resolved 3-D imaging of live cells' organelles from bright-field photon transmission micrographs.

Author information

1
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.
2
Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
3
Princeton University, Princeton, New Jersey 08544, USA.
4
Department of Chemistry, Stanford University, Stanford, California 94305, USA.

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.

KEYWORDS:

3D; Bright-field microscopy; Light transmission; Live cell imaging; Super-resolution

PMID:
28364682
DOI:
10.1016/j.ultramic.2017.03.018
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center