Effect of heparin as an anticoagulant on measurements of the erythrocyte genome size using flow cytometry in bony fishes of different ploidy

Halačka, Karel; Janko, Karel; Fedorčák, Jakub; Koščo, Ján; Vetešník, Lukáš

doi:10.2754/avb202392020171

Acta Veterinaria > Archive > 2023, Vol. 92 > Issue 2 > Effect of heparin as an anticoagulant on…

Acta Vet. Brno 2023, 92: 171-179

https://doi.org/10.2754/avb202392020171

Effect of heparin as an anticoagulant on measurements of the erythrocyte genome size using flow cytometry in bony fishes of different ploidy

Karel Halačka^1,2, Karel Janko^1,3, Jakub Fedorčák⁴, Ján Koščo⁴, Lukáš Vetešník²

¹Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Liběchov, Czech Republic
²Czech Academy of Sciences, Institute of Vertebrate Biology, Brno, Czech Republic
³University of Ostrava, Department of Biology and Ecology, Ostrava, Czech Republic
⁴University of Prešov in Prešov, Faculty of Humanities and Natural Sciences, Department of Ecology, Prešov, Slovakia

Received May 10, 2021
Accepted May 4, 2023

The aim of the study was to verify whether the use of heparin as an anticoagulant can affect the results of genome size measurements using a flow cytometer. The values obtained by measuring blood samples with different concentrations of heparin were compared. The differences observed were highly significant, suggesting that the results of previous comparative studies, or those determining individual polyploidy, may need reinterpretation. We anticipate that our findings will increase the accuracy of flow cytometric analysis and will be useful for further surveys of genome size undertaken using blood samples.

Keywords

DAPI, propidium iodide, DNA content, C-value, fish.

Funding

We thank Dr. Vladislav Kolarčik for help with the PARTEC CyFlow®ML analysis of the control blood samples and David Lee McLean for help with the English language correction. This study was supported through the Grant Agency of the Czech Republic, Grant Nos. GACR 19-21552S. We are also grateful to the contribution of the Slovak Research and Development Agency under the project APVV SK-AT-20-0009 and Scientific Grant Agency of the Slovak Republic under the grants VEGA 1/0553/23.