Fixation of cell-bound antibody in the membrane immunofluorescence test
| Autor: | Peter Biberfeld, Zelma Molnar, Gunnel Biberfeld, Astrid Fagraeus |
|---|---|
| Rok vydání: | 1974 |
| Předmět: |
Cell Membrane Permeability
Immunology Fluorescent Antibody Technique Immunofluorescence Antigen-Antibody Reactions Cell membrane chemistry.chemical_compound Antigen Formaldehyde Methods medicine Animals Humans Immunology and Allergy Fluorometry Horses Lymphocytes Fixative Fixation (histology) Sheep medicine.diagnostic_test biology Chemistry Immune Sera Cell Membrane Histological Techniques Osmium medicine.anatomical_structure Microscopy Fluorescence Osmium tetroxide Biochemistry Glutaral Immunologic Techniques Biophysics biology.protein Rabbits Glutaraldehyde Lymphocyte Culture Test Mixed Antibody |
| Zdroj: | Journal of Immunological Methods. 4:135-148 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/0022-1759(74)90056-8 |
| Popis: | Fixative treatment to stabilize cell-bound antibody was described and applied in quantitative and qualitative studies of cell membrane antigens by direct and indirect immunofluorescence. Of the several fixatives tested, namely p -formaldehyde (PFA), glutaraldehyde (GA) acetone and osmium tetroxide, PFA appeared to have several advantages compared to the other fixatives. Fixation with PFA under specified conditions did not diminish the fluorescence intensity of cells stained by direct or indirect immunofluorescence and decreased only slightly the reaction of anti-immunoglobulin with cell-bound fixed antibody. Redistribution phenomena as ‘capping’ and dot-formation were abolished or greatly reduced by PFA fixation of the cell-bound antibody in the indirect membrane fluorescence test. The characteristic hand-mirror configuration, due to uropod formation of stimulated lymphocytes was retained if the cells were PFA-fixed at 37°C. After PFA-fixation of stimulated lymphocytes which had been incubated under conditions favouring redistribution of cell-bound antibody it was evident that ‘capping’ was confined to the side of the cells which formed the uropod. |
| Databáze: | OpenAIRE |
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