Establishment of HEp‐2 cell preparation for automated analysis of ANA fluorescence pattern
- AbstractBackgroundIdentification of antinuclear antibodies (ANAs) has large clinical importance for the assessment of autoimmune diseases. HEp‐2 cell preparations on microscopic slides are commonly used as antigenic substrate. Methods used for cell preparation are important for ANA pattern analysis; however, these methods differ widely and are mostly not specified.MethodsHEp‐2 cells were fixed using acetic acid‐ethanol, methanol‐acetone, acetone, formaldehyde, paraformaldehyde, or glutaraldehyde. Morphological analysis was done after haematoxylin‐eosin staining and DAPI‐staining of cell nuclei.ResultsThe results demonstrate a high variability of cell and nuclear morphology depending on the used fixatives. Aldehyde fixatives conserved the cell structures best, acetone fixatives revealed remarkable changes.ConclusionsAfter selecting appropriate fixation procedures to preserve nuclear structures further experiments are necessary to find out which fixation procedure preserves the disease‐linked antigens the best way and are, therefore,AbstractBackgroundIdentification of antinuclear antibodies (ANAs) has large clinical importance for the assessment of autoimmune diseases. HEp‐2 cell preparations on microscopic slides are commonly used as antigenic substrate. Methods used for cell preparation are important for ANA pattern analysis; however, these methods differ widely and are mostly not specified.MethodsHEp‐2 cells were fixed using acetic acid‐ethanol, methanol‐acetone, acetone, formaldehyde, paraformaldehyde, or glutaraldehyde. Morphological analysis was done after haematoxylin‐eosin staining and DAPI‐staining of cell nuclei.ResultsThe results demonstrate a high variability of cell and nuclear morphology depending on the used fixatives. Aldehyde fixatives conserved the cell structures best, acetone fixatives revealed remarkable changes.ConclusionsAfter selecting appropriate fixation procedures to preserve nuclear structures further experiments are necessary to find out which fixation procedure preserves the disease‐linked antigens the best way and are, therefore, suitable to be used in ANA‐testing of AABs. © 2006 International Society for Analytical Cytology…
MetadatenAuthor: | Daniel Hahm, Ursula Anderer |
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DOI: | https://doi.org/10.1002/cyto.a.20223 |
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ISSN: | 1552-4922 |
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Title of the source (English): | Cytometry. Part A |
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Document Type: | Scientific journal article peer-reviewed |
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Language: | English |
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Year of publication: | 2006 |
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Tag: | Antinuclear antibodies; Autoimmune diagnostic; Cell Biology; HEp-2 cells; Histology; Pathology and Forensic Medicine |
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Volume/Year: | 69A |
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Issue number: | 3 |
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First Page: | 178 |
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Last Page: | 181 |
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Fundername (not EU): | Bundesministerium für Wirtschaft und Technologie |
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Project number (not EU): | 03WKR05C |
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Way of publication: | Open Access |
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Faculty/Chair: | Fakultät 2 Umwelt und Naturwissenschaften / FG Zellbiologie |
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