Analysis of living cell 3D inner structures from high-resolution bright-field microscopy

R. Rychtáriková, T. Náhlík, D. Štys

University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Institute of Complex Systems, Zámek 136, 373 33 Nové Hrady, Czech Republic

Knowledge of composition of biochemical structures (from organelles, bio-molecular assembles down to unique biomolecules) is important for understanding of structural-functional relationships inside a living cell and recognition its physiological state. The best imaging technique is provided by a classical microscopic technique without usage any contrast methods – bright-field light transmission microscopy. Provided we use an unlabelled, undyed cell, we obtain its most real image.

With regards to physical and physico-chemical process, which occur during passing light through a living cell, we developed a mathematical approach (Point Divergence Gain) for segmentation and analysis of three-dimensional structures down to size of a camera chip (x,y-coordinates) and scanning z-step from their light diffraction and emission patterns – object spread functions – as well as tracking organelles captured in a z-stack of micrographs [1].

Our results show necessity to consider the technical limitations of microscopy (i.e., precision of optics, image registration, size of scanning z-step and a camera pixel, speed of image acquisition and storage) in future building-up of a new microscope and relevant software. 

1. R. Rychtáriková, T. Náhlík, R. Smaha, J. Urban, D. Štys Jr., P. Císař, D. Štys, in ISCS 2014:Interdisciplinary Symposium on Complex Systems (Emergence, Complexity and Computation 14), edited by A. Sanayei, O.E. Rössler, I. Zelinka (Switzerland: Springer), 2014, pp. 261-267.

This work was financially supported by Postdok JU CZ.1.07/2.3.00/30.0006, by the GAJU 134/2013/Z, and by the Ministry of Education, Youth and Sports of the Czech Republic projects CENAKVA (No. CZ.1.05/2.1.00/01.0024) and CENAKVA II (No. LO1205 under the NPU I program).