[doi: 10.5505/2017ichc.PP-62]Potential difficulties in transmission electron microscope observations of respiratory cilia axonemal complexOya Evirgen, Esra Erdemli, Emine Nazli Hayirli, Derya OzdemirOBJECTIVES: Primary ciliary dyskinesia(PCD) is a rare and genetically heterogenous group of recessive disorders with various ultrastructural defects of axonem and impaired ciliary motility resulting in chronic upper and lower respiratory tract disease, fertility problems and organ laterality defects. Transmission electron microscopic (TEM) analysis of cilia is still an important part of diagnosis and optimal fixation, processing and sectioning of the biopsies are neccessary to study the internal axonemal structure of cilia. In this study we try to check up the sample preperation and sectioning steps performed in our TEM laboratory. MATERIALS-METHODS: Biopsies were obtained from either bronchial or nasal mucosa. The removed tissue sample and the pellet of lavage solution after centrifugation were resuspend and fixed in fresh 2,5% glutaraldehyde and processed as usual for ultrastructural analysis. The semi-thin sections stained with toluidine blue examined under light microscopy. After staining with uranyl acetate/lead citrate the different thickness of (85nm,100nm) ultra-thin sections were observed under LEO 906 E transmission electron microscope. RESULTS: In semi-thin sections of nasal and bronchial biopsies showed normal ciliated respiratory epithelial cells and goblet cells. And some of semi-thin sections ciliated epithelial cells can not be observed. The ultra-thin sections showed transverse, oblique and longitudinal sections of cilia. Most of them have normal axonemal pattern with 9+2 configuration while in a few cilia showed microtubular disorganization. CONCLUSION: TEM is often a useful tool in defining the ultrastructural defects of axonem and has been used for the diagnosis of PCD. But it can not be helpful to identify the PCD variants with normal ultrastructure. So recently new methods for diagnosis such as immunofluorescence analysis has been used to investigate the subcellular localization of the ciliary proteins. For TEM examination the biopsy site(nasal or bronchial), experience of clinician performing biopsy and accurate patient selection is important for a feasible ultrastructural analysis. Also an optimal specimen processing steps, achivement of sufficient number of high quality transverse sections of cilia is very important to study the internal axonemal structure of respiratory cilia. |