caa a22 4500 465774121 CHVBK 20180323111937.0 cr unu---uuuuu 170327e19900101xx s 000 0 eng 10.1007/BF00412737 doi (NATIONALLICENCE)springer-10.1007/BF00412737 Three-dimensional visualization of human Langerhans' cells using confocal scanning laser microscopy [Elektronische Daten] [A. Scheynius, P. Lundahl] Summary: Most techniques used to visualize cells in tissues are accompanied by some distortion of the tissue due to fixation and sectioning. We here present a technique involving "optical sectioning” and three-dimensional reconstruction of fluorescence-labelled Langerhans' cells in human skin that avoids such problems. The instrument used to study the specimens was a PHOIBOS Confocal Scanning Laser Microscope (CSLM) built around a Zeiss Universal microscope. A software package for three-dimensional reconstructions of confocal sections was employed for calculations in a Hewlett-Packard HP9000-350 minicomputer running UNIX. Normal human skin obtained at plastic surgery or skin biopsy specimens from allergic, irritant, or control patch test reactions was studied. Epidermis separated from dermis by incubation in thermolysin or thick (25 μm) cryostat sections were incubated with fluorescein-isothiocyanate labelled mouse monoclonal antibodies directed against HLA-DR. To reduce the fading of immunofluorescence during microscopy, the specimens were mounted with glycerol in PBS containing p-phenylenediamine. The results indicate that CSLM is a powerful tool for investigations on Langerhans' cells in different conditions of the skin allowing a three-dimensional view of cells in unfixed or acetone-fixed preparations at the light microscope level. Springer-Verlag, 1990 Langerhans' cells nationallicence Human skin nationallicence Confocal scanning laser microscope nationallicence 3-D reconstruction nationallicence Scheynius A. Department of Clinical Immunology, Karolinska Institute, Stockholm, Sweden aut Lundahl P. Sarastro AB, Stockholm, Sweden aut Archives of Dermatological Research Springer-Verlag 281/8(1990-01-01), 521-525 0340-3696 281:8<521 1990 281 403 https://doi.org/10.1007/BF00412737 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00412737 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Scheynius A. Department of Clinical Immunology, Karolinska Institute, Stockholm, Sweden aut NATIONALLICENCE 700 1- Lundahl P. Sarastro AB, Stockholm, Sweden aut NATIONALLICENCE 773 0- Archives of Dermatological Research Springer-Verlag 281/8(1990-01-01), 521-525 0340-3696 281:8<521 1990 281 403 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer