Acta Vet. Brno 2021, 90: 225-231

Comparison of cryoprotective methods for histological examination of rat and porcine lung tissue

Rea Jarošová1, Petra Ondráčková2, Zdeněk Patočka3, Zbyšek Sládek1

1Mendel University in Brno, Faculty of AgriSciences, Department of Morphology, Physiology and Animal Genetics, Brno, Czech Republic
2Veterinary Research Institute, Department of Immunology, Brno, Czech Republic
3Mendel University in Brno, Faculty of Forestry and Wood Technology, Department of Forest Management and Applied Geoinformatics, Brno, Czech Republic

Received August 13, 2020
Accepted May 26, 2021

Proper histological evaluation of lung tissue and accurate antigen detection by immunodetection techniques require histological tissue processing to preserve tissue reactivity and open alveolar spaces. In this study, we focused on testing and comparing different procedures of tissue cryopreservation. Sucrose or Tissue Tek were used with several methods of freezing samples by supercooled liquids and liquid nitrogen. Changes in tissue caused during the freezing of samples and the effect of cryoprotectants on the tissue were recorded. Rat and porcine pulmonary tissues were used in this experiment. This study aimed to optimize the process of lung cryopreservation with emphasis on enabling proper anatomical evaluation and preserving a high tissue immunoreactivity. The best results were obtained by inflating pulmonary parenchyma with a 1 : 1 mixture of O.C.T. with phosphate buffered saline (PBS) frozen in supercooled n-heptane placed on dry ice. Pulmonary tissue prepared in this way enables to perform proper histological evaluation and to detect target molecules by immunohistochemical analysis.


This research project AF-IGA-IP-2018/021 was financially supported by the Internal Grant Agency of Mendel University in Brno and the Ministry of Agriculture of the Czech Republic (RO 0519).


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