Zellforschung
Notizen zu den Entwicklungswegen der Mikroskopie, Histologie und Methoden für die Herstellung klassischer Färbelösungen und Reagenzien. Unser Blick richtet sich auf immunhistologische und molekularspezifische Prinzipien zur Darstellung von Biomarkern. Anwendungsbeispiele unterstreichen ihre Nützlichkeit für das Studium der zellulären Differenzierung in Ontogenese, Krebsforschung und Histopathologie.
Microskopie, Histologie
Innovative Verfahren ermöglichen Einblicke in strukturelle Details. Mit komplexen Methoden wird eine hohe Spezifität erreicht.
- Immunohistology: an introduction to selective cell labeling as an essential element in the study of structure-function relationship
- The microscope as a scientific tool
- Fluorescence and fluorescence microscopy
- Histological staining techniques
- Dyes, stains, and special probes in histology
- Immuno-enzyme techniques in cytochemistry
- General aspects in tissue preparation
- Fixation of biological specimens
- Tissue dehydration, embedment
- Cryostat, cryotechniques
- Microtomy of tissue specimens, collection of sections
Antikörper, Markermoleküle, Konjugate
Immunhistologische Zellmarkierungen erfordern spezifische Antikörper. Die Detektion der Antigen-Antikörperreaktionen im Gewebe basiert auf der Verwendung von Markermolekülen mit definierten physiko-chemischen Eigenschaften. Geeignete Markiermoleküle sind beispielsweise Fluorochrome und Enzyme als Reporter der selektiven Bindungsreaktion.
- Antibody molecules and the antigen-antibody interaction
- Immunization, immune sera and the production of antibodies
- Specificity of antibodies
- Purification of antibodies by use of biochemical and immunochemical techniques
- Immunohistological marker molecules
- Fluorochromes in selective cell analysis
- Enzymes as marker molecules
- Markers for immuno-electron microscopy
- Ferritin as electron dense label
- Gold labelled reagents as marker system
- Preparation of conjugates
Selektive Zellfärbung
Biomarker können in histologischen Präparaten mit direkten oder indirekten Markierungs-Prinzipien dargestellt werden. Das jeweils geignete Format wird experimentell aus einer Vielzahl verfügbarer Möglichkeiten ausgewählt. In der Regel muß das Testformat an die spezielle Anforderung angepasst werden. Dies betrifft sowohl das zu untersuchende Gewebe als auch die nachzuweisende Molekülstruktur.
- Cell staining with direct and indirect assay formats
- Considerations prior to immuno-staining
- Retrieval of antigenic determinants
- Inhibition of endogenous enzyme activity
- Staining enhancement, signal amplification
- Lectins and nucleic acids as molecular probes
- Immuno-staining of paraffin embedded tissue sections
- Immuno-staining of resin embedded tissue sections
- Preembedding immuno-staining of specimens for electron microscopy
- Immunofluorescence staining of cryostat sections
- Immunofluorescence assay for autoantibodies to nuclear antigens (ANA) on HEp-2-cells
- Specificity and standardization of immunohistology
- Practical aspects in quality control of immunohistology
Reagenzien, Methoden
Auswahl von häufig verwendeten Reagenzien und Lösungen für Immunhistologie, Zytochemie und Biochemie. Die Reinheit der Reagenzien ist entscheidend für die Qualität der erzielten Ergebnisse.
- Fixatives
- Buffer solutions
- Washing solutions
- Blocking solutions
- Incubation and antibody dilution buffers
- Enzyme cytochemical substrate solutions
- Natural and synthetic dyes in histology
- Conjugation of antibodies by use of glutaraldehyde
- Protein labeling with biotin derivatives
- Preparation of soluble PAP complexes
- Measurement of peroxidase (HRP) enzyme activity in conjugates
- Measurement of glucose oxidase (GOD) enzyme activity in conjugates
- Dehydration and paraffin embedding of tissues
- Dehydration an resin embedment of tissues
- Silane conditioning of glass slides
- Elimination of mercury salts from tissue sections
- Peroxidase (HRP) cytochemistry
- Glucose oxidase (GOD) cytochemistry
- Haematoxylin staining methods
- Carmine staining methods
- Gallocyanin-chrome alum method
- Methyl green counterstaining
- Toluidine blue staining
- Romanowsky_Giemsa staining
- Periodic acid-Schiff (PAS) reaction
- Staining with Alcian blue at pH 2.5
- Azure-methylene blue staining of sections from resin embedded tissues
- Acronyms and terms in cell research and microscopy
Anwendungsbeispiele aus der Zellforschung
Beispiele histologischer Untersuchungen von Biomarkern in Gewebeschnitten für die Lokalisierung von onkofetalen Proteinen (Alpha-Fetoprotein), Enzymen und Glykokonjugaten in experimentellen Modellen und in histopathologischen Präparaten.
- Cellular fluctuations, alpha-fetoprotein expression, and the question of stem cells in experimental liver regeneration and hepatocarcinogenesis
- Hepatic progenitor cells, stem cells, and AFP expression in models of liver injury
- Immuno-electron microscopy of alpha-1-fetoprotein during normal development of rat hepatocytes
- Ultrastructural detection of alpha1-fetoprotein in hepatomas by use of peroxidase-labelled antibodies
- Focal elevation of liver microsomal epoxide hydrolase in early preneoplastic stages and its behaviour in the further course of hepatocarcinogenesis
- Regulation and expression of four cytochrome cytochromes P-450 isoenzymes, NADPH-reductase, the glutathione transferases B and C and microsomal epoxide hydrolase in preneoplastic and neoplastic lesions in rat liver
- Expression and inducibility of drug-metabolizing enzymes in preneoplastic and neoplastic lesions of rat liver during nitrosamine-induced hepatocarcinogenesis
- Alpha-fetoprotein: cellular origin of a biological marker in rat liver under various experimental conditions
- Characterization, localization and regulation of a novel phenobarbital-inducible form of cytochrome P450-isoenzymes, NAPH P450-reductase, glutathione transferases and microsomal epoxide hydrolas
- Correlation of histology and alpha-1-fetoprotein resurgence in rat liver after experimental injury by galactosamine
- Hepatic progenitor cells, stem cells, and the biliary epithelium as transit compartment in models of liver injury
- Localization of alpha-1-fetoprotein and DNA-synthesis in liver cell populations during experimental hepatocarcinogenesis in rats
- Immunoperoxidase labelling of alpha-1-fetoprotein (AFP) in normal and regenerating livers of a low and a high AFP producing mouse strain
- Lectin-peroxidase conjugates in histopathology of gastrointestinal mucosa
- Cellular localization of lectin affinity in tissue sections of normal human duodenum
- Comparative study of procedures for histological detection of lectin binding by use of Griffonia simplicifolia agglutinin I and gastrointestinal mucosa of the rat
- Glucose oxidase as label in histological immunoassays with enzyme-amplification in a two-step technique: coimmobilized horseradish peroxidase as secondary system enzyme for chromogen oxidation
- Resin embedment of organs and postembedment localization of antigens by immunoperoxidase methods
- Ultrastructural immunoperoxidase cytochemistry
- Purification of mouse alpha-1-fetoprotein and preparation of specific peroxidase conjugates for its cellular localization
- Comparative study of the techniques for ultrastructural localization of antienzyme antibodies
- Comparative study for ultrastructural localization of intracellular immunoglobulins using peroxidase conjugates
- Localisation intracellulaire d’anticorps à l’aide de la glucose oxidase comme antigène et marqueur
- Immunohistochemical studies on human gastric mucosa. Procedures for routine demonstration of gastric proteins by immunoenzyme techniques
- Glucose oxidase as an antigen marker for light and electron microscopic studies
- Diaminobenzidine cytochemistry in cryoultramicrotomy for the detection of peroxidases
- Cross-linked albumin as supporting matrix in ultrathin cryo microtomy
- Immunoperoxidase technique in cryoultramicrotomy for the detection of simian adenovirus in cell nuclei
- Correlation of cell division and specific protein production during the course of lymphoid cell differentiation
- Lymphocyte differentiation and antibody synthesis in the secondary immune response of peroxidase stimulated lymph nodes of rat
- Cellular differentiation and antibody localization during the primary immune response in peroxidase stimulated lymph nodes of rat
- Immunofluorescent histological demonstration of the antigenic gastric mucosal esterase VIA
in the superficial epithelium of human fundic gastric mucosa - Species specificity of the human gastric mucosa esterase VIA. I. Comparative studies on
gastric mucosa extracts of rabbit, guinea pig, rat and men
Photo-Archiv Immunhistologie
Mikroskopisch-histologischer Nachweis von biochemisch definierten Zellstrukturen mit mono- oder polyklonalen Antikörpern für die Detektion von reaktiven Molekülen. Sinngemäß kann diese Technik auch auf andere Strukturen mit unterschiedlichem Trägermaterial für den spezifischen Nachweis molekularer Eigenschaften übertragen werden. Es können qualitative und quantitative Liganden-Assays mit variablen Detektionsverfahren entwickelt werden. Sowohl direkte als auch indirekte Nachweisformen sind möglich.
- Principles and technical approaches for the detection of molecular staining in immunohistology and other ligand binding assays
- Expression of AFP in hepatic regeneration and hepatocarcinogenesis
- Cellular AFP in normal liver
- Serum AFP levels after acute injury and during hepatocarcinogenesis
- AFP expression by hepatocytes and biliary epithelial cells (oval-cell proliferation) during liver regeneration and chemical hepatocarcinogenesis
- AFP resurgence and histological lesions in late stages of chemical hepato-carcinogenesis and during hepatoma development
- Selection of organ-type antigens in normal and diseased tissues
- Lymphocyte differentiation and detection of anti-enzyme antibody synthesis in lymph node cells after peroxidase (HRP) immunization
- Lectins as specific ligands for the study of glycoconjugates in cell research and histopathology
Photo-Archiv Methoden und Reagenzien
Material und Laborausstattung für die Aufbereitung von Antigenen, Antikörpern und Antikörper-Enzym Konjugaten. Bearbeitung von Gewebeproben für die Mikroskopie.
Referenzen
Publikationen zu Mikroskopie, histologischen Techniken und methoden-relevanten Arbeitsgebieten.