Spectroscopic Approach to Correction and Visualisation of Bright-Field Light Transmission Microscopy Biological Data
Ganna Platonova,
Dalibor Štys,
Pavel Souček,
Kirill Lonhus,
Jan Valenta,
Renata Rychtáriková
Affiliations
Ganna Platonova
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, Institute of Complex Systems, University of South Bohemia in České Budějovice, Zámek 136, 373 33 Nové Hrady, Czech Republic
Dalibor Štys
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, Institute of Complex Systems, University of South Bohemia in České Budějovice, Zámek 136, 373 33 Nové Hrady, Czech Republic
Pavel Souček
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, Institute of Complex Systems, University of South Bohemia in České Budějovice, Zámek 136, 373 33 Nové Hrady, Czech Republic
Kirill Lonhus
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, Institute of Complex Systems, University of South Bohemia in České Budějovice, Zámek 136, 373 33 Nové Hrady, Czech Republic
Jan Valenta
Faculty of Mathematics and Physics, Charles University, Ke Karlovu 3, 121 16 Prague, Czech Republic
Renata Rychtáriková
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, Institute of Complex Systems, University of South Bohemia in České Budějovice, Zámek 136, 373 33 Nové Hrady, Czech Republic
The most realistic information about a transparent sample such as a live cell can be obtained using bright-field light microscopy. Under high-intensity pulsing LED illumination, we captured a primary 12-bit-per-channel (bpc) response from an observed sample using a bright-field microscope equipped with a high-resolution (4872 × 3248) image sensor. In order to suppress data distortions originating from the light interactions with elements in the optical path, poor sensor reproduction (geometrical defects of the camera sensor and some peculiarities of sensor sensitivity), we propose a spectroscopic approach for the correction of these uncompressed 12 bpc data by simultaneous calibration of all parts of the experimental arrangement. Moreover, the final intensities of the corrected images are proportional to the photon fluxes detected by a camera sensor. It can be visualized in 8 bpc intensity depth after the Least Information Loss compression.
