The lysosomal copper concentration in the liver in primary biliary cirrhosis.

The copper concentration in the lysosomes of periportal hepatocytes of nine patients with primary biliary cirrhosis was determined by X-ray microanalysis in ultrathin sections, the chlorine concentration of the embedding medium being taken as internal standard. A positive correlation was found between the lysosomal and the total liver copper concentration. No correlation was found between the lysosomal copper concentration and the grade of orcein staining for lysosomal copper-associated protein in paraffin sections. It is concluded that in contrast to the grade of orcein staining, the lysosomal copper concentration is a reliable measure of the total liver copper concentration in primary biliary cirrhosis.

Methods in laboratory investigation. Immunoelectron microscopic methods for demonstration of antigens on normal human melanocytes and other epidermal cells.

Different procedures for fixation and processing were evaluated in order to examine the antigenic profile of melanocytes and other epidermal cells for immunoelectron microscopy. For this purpose the monoclonal antibodies anti-HLA-A, B, C, anti-HLA-DR, anti-T6, and the melanoma-associated monoclonal antibody NKI /C-3 were used as markers. Fixation with periodate-lysine-paraformaldehyde yielded better antigenic and ultrastructural preservation than 3% paraformaldehyde or picric acid-paraformaldehyde did. Skin was further processed by five different methods: (a) 15-micron frozen sections; (b) 75-micron agar-embedded, tissue chopper sections; (c) 15-micron polyethylene glycol-embedded sections; (d) epidermal cells in suspension; and (e) epoxy sections (postembedding staining) were prepared for the immunoperoxidase procedure. Antigenicity was best preserved in the cell suspension method and somewhat less, but with a similar staining distribution, with the first three methods. Staining with the polyethylene glycol-embedded sections was only achieved if they were left free-floating in buffer; no staining was observed when the sections were mounted on glass slides and left to dry overnight at 37 degrees C. Epidermal cells remained unreactive in the postembedding method, even after etching. Ultrastructural preservation of the agar-embedded sections and the cells in suspension was superior to the other preembedding methods. Melanocytes mostly showed moderate staining for HLA-A, B, C and slight staining for the antigen that is recognized by NKI /C-3. The latter was further demonstrated on Langerhans cells and indeterminate cells which also expressed HLA-A, B, C, T6, and HLA-DR antigens.

Chemodectoma (paraganglioma) of the larynx.

Two cases of chemodectoma of the larynx, 1 benign and 1 malignant, are reported. Initially both tumours caused diagnostic problems but the diagnosis could be confirmed by silver staining and electron microscopy. So far only 25 cases of laryngeal chemodectoma have been reported in the literature. Clinically, the diagnosis should be considered when a persistent, smooth, non-inflammatory intralaryngeal swelling, particularly with a reddish-blue colour, is encountered. A final histological diagnosis can only be made with additional histochemical techniques and electron microscopy.

Ultrastructure of normal epithelial cells in Papanicolaou-stained cervical smears. An application of a modified open-face embedding technique for transmission electron microscopy.

The results of an open-face embedding technique applied to Papanicolaou-stained uterine cervical smears are presented. By this technique, cells are smeared on a plastic sheet, fixed in 1.5% buffered glutaraldehyde and stained by the Papanicolaou method; areas of special interest are then selected by light microscopy for electron microscopic observations. Thus, a comparison is possible between the light microscopic characteristics of cells in smear preparations and their ultrastructural counterparts. The good ultrastructural preservation allowed detailed study of nuclei and cytoplasm. The endocervical cells had intact nuclei with nuclear envelope, Golgi apparatus, well-preserved mitochondria and two types of secretory material. The superficial and intermediate squamous cells showed loss of desmosomes; dense, fine cytoplasmic fibrils; small, fatty droplets; and degenerated nuclei with loss of the nuclear envelope. Compared with ultrastructural studies on cervical tissues, the intermediate cells in cervical smears were more degenerated, presumably because predominantly degenerated intermediate cells exfoliate. This study indicates that the light microscopic impression of the nuclear appearance is misleading: in the routine smear the nuclei of superficial and intermediate cells may look "intact" and "well preserved" whereas ultrastructurally the nuclei are degenerated.

Legionnaires' disease: case report with special emphasis on electron microscopy and potential risk of infection at autopsy.

A case of Legionnaires' disease (LD) is presented in which the diagnosis was made initially by EM and later confirmed by specific immunofluorescence of formalin-fixed, paraffin-embedded lung tissue. The possibility of an infection with Legionella pneumophila during autopsy is suggested by the fact that the pathologist who performed the post-mortem examination developed signs and symptoms of an upper respiratory tract infection 2 weeks afterwards. Approximately 4 weeks after the onset of his illness a significant antibody titre against L. pneumophila was found in his serum, whereas 6 months later the antibody titre was insignificant.