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Histological and histochemical methods used in study of development of internal male sex organs
 VÁVROVÁ, Martina
Presented thesis devotes to comparative study of usability of fixation agents and colouring methods for the study of histology and histochemistry of Culex pipiens s.l. mosquito males. The study was carried out in males of Culex pipiens quinquefasciatus Say, which is a significant transmitter of filariasis. Histological and histochemistry studies of the structure of the whole abdomens or prepared testicles were carried out on the material fixed by fixation agents Carnoy and Bouin of Dubosque-Brasil modification. Fixed tissue was cast in paraplast or resin. Histological sections were done by means of automatic microtome Leica and half-thin sections by means of ultra microtone Reichert. Section preparations were coloured by Mayer or Harris haematoxylin or by colouring method according to Mallory. Semi-thin sections from materials cast in resin were coloured by toluidine blue. Fixation agent Bouin, Dubosque-Brasil modification proved as more convenient than Carnoy, either for the preparation of paraffin sections or for fabrication of half-thin sections from tissues cast in resin. The next advantage of this fixation is that fixed tissue can be stored for unlimited period of time without over fixation of the tissue as it is in the case of fixation by Carnoy. For general histological preparations Mallory colouring is apparently the most convenient from the used methods; it provides better orientation in coloured tissue thanks to the whole spectrum of colours-pink to red colouring of cell nuclei, blue colouring of tissue and secretions. In case of colouring of male genital organs all used colourings (Mallory, Harris and Mayer haematoxylin) seem to be comparable. Casting in resin with connection to colouring by toluidine blue (also histochemistry proof for carbohydrates) has shown to be suitable for visualisation of finer structures, in male genital organs particular developmental phases of male sex cells were distinguished.
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