Use of periodate-lysine-paraformaldehyde for the fixation of multiple antigens in human skin biopsies
| Autor: | L. Domenici, Laura Pieri, C. Sassoli, Paolo Romagnoli |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Keratinocytes
Tissue Fixation Histology Langerhans cell Biopsy Biophysics membrane antigens bromo-deoxyuridine chemical and pharmacologic phenomena Human skin tyrosinase Biology PLP Acetone Immunoenzyme Techniques Fixatives chemistry.chemical_compound Immunolabeling Antigen Formaldehyde Istochimica immunoistochimica microscopia elettronica fissazione tecnica istologica medicine Humans Langerhans cells Antigens Microscopy Immunoelectron lcsh:QH301-705.5 Fixative Skin Monophenol Monooxygenase Lysine Cell Cycle Periodic Acid cytokeratins Cell Biology Immunohistochemistry Molecular biology Microscopy Electron medicine.anatomical_structure lcsh:Biology (General) Biochemistry chemistry Indicators and Reagents Glutaraldehyde cryosections Keratinocyte |
| Zdroj: | European Journal of Histochemistry, Pp 365-75 (2010) |
| ISSN: | 2038-8306 1121-760X |
| DOI: | 10.4081/1749 |
| Popis: | Periodate-lysine-paraformaldehyde (PLP) has been proposed as a fixative for glycoprotein antigens which should stabilize periodate oxidized polysaccharide chains through lysine mediated crosslinks, either directly or by the intermediation of formaldehyde. In spite of premises and attempts reported in the literature, this fixative has never become popular for the study of membrane antigens of immune system cells, which leads to doubts on its real efficacy. We have addressed this issue in biopsies of human skin and found that PLP followed by cryoprotection with 30% sucrose and cryosectioning, or PLP fixation of isolated epidermal sheets, consistently provided for good preservation of morphology and intense labeling of major histocompatibility complex class II molecules, CD1a, CD4, CD8, E-cadherin, cytokeratins in general, cytokeratin-18 in particular, and bromodeoxyuridine, incorporated by cycling cells in vitro, and for the demonstration of tyrosinase enzyme activity. PLP-fixed, osmicated and epon-embedded epidermal sheets proved as good as sheets fixed 365 with a mixture of formaldehyde and glutaraldehyde for electron microscopic morphological analysis. Also, these sheets were amenable to immunoperoxidase staining of Langerhans cell membrane antigen CD1a and keratinocyte membrane antigen E-cadherin before being osmicated and prepared for electron microscopy. In a parallel paper, we had also shown that oral mucosa biopsies fixed in PLP showed good morphology and immunolabeling of CD54, CD80, CD83 and CD86. Therefore, we conclude that PLP can be proposed as a multi-task fixative for light and electron microscopic analysis of membrane, cytoplasmic and nuclear antigens of immune system cells and keratinocytes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|