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Morfocytometric investigation of haemocytes in comparative hematology
ŠUBJAK, David
Standard haematological procedures in preclinical toxicity studies are mostly carried out on mammals, more recent haematotoxicological models represent invertebrates because they can be used in the screening of myelotoxicity. In the present study, we examined haemocytes of Culex pipiens as a new potential alternative haematological in vivo model. Preparations for microscopic analysis were made from the hemolymph stained by Pappenheim's method for overstriking morphology and by toluidin blue and AgNOR for overstriking nucleoles. Microscopic photos were processed by image analysis and size of cells, nuclei and nucleoli were evaluated. Six types of haemocytes were found: granulocytes, prohemocytes, spherulocytes, plasmatocytes, adipohemocytes and oenocytes. Thanks to image analysis we made a morfometric profile of each type of haemocytes and set the nucleolar coefficient. We described significant differences among prohemocytes, plasmocytes and oenocytes size. On the grounds of nucleoles size we divided haemocytes to next two significant groups. Image analysis seems to be very helpful for distinction among various haemocyte lines but we cant use it for generally separation without personal control thanks to presence of statistically significant findings for only some comparisons cell types.
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Cytochemická vyšetření akvitivity nukleolů po podání genisteinu na modelovém objektu
PICMONOVÁ, Veronika
This thesis deals with exploration of the effect of genistein on proteosynthetic activity of nucleoli by observing changes in their morphology. The effect of genistein on the function of nucleoli was observed in hemocytes of the experimental animal Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae). 4 types of hemocytes were found in the hemolymph of S. littoralis (Boisduval): granulocytes, prohemocytes, plasmatocytes and spherulocytes. The effect of genistein on individual types and concentration of hemocytes is not clear. Genistein was applied in doses of 0,135 mg and 0,54 mg per larva, and subsequently changes in morphology, number and size of nucleoli were examined upon 8 and 24 hours. Furthermore, changes in morphology of the hemocytes were examined, as well as their concentration using toluidine blue staining, silver staining, panoptic staining and using the Türk{\crq}s solution. For the purpose of changes assessment, the material was processed using an optical microscope and the computer software Nis Element AR 2.30. In the course of observation, changes in the number of nucleoli were found upon application of genistein. The number of compact nucleoli and micronucleoli increased. On the contrary, the number of ring-shaped nucleoli decreased. The area of nucleoli increased upon application of lower dosage of genistein (0,135 mg); however, upon action of the higher dose (0,54 mg), it decreased, on the contrary. Nucleolar index values remained without rather significant changes. In the biomodel used, genistein evoked changes which can be viewed as immune responses from the viewpoint of hematotoxicology.
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