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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 Cytologyshow moreshow less

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Metadaten
Author: Daniel Hahm, Ursula Anderer
DOI:https://doi.org/10.1002/cyto.a.20223
ISSN:1552-4922
Title of the source (English):Cytometry. Part A
Document Type:Scientific journal article peer-reviewed
Language:English
Year of publication:2006
Tag:Antinuclear antibodies; Autoimmune diagnostic; Cell Biology; HEp-2 cells; Histology; Pathology and Forensic Medicine
Volume/Year:69A
Issue number:3
First Page:178
Last Page:181
Fundername (not EU):Bundesministerium für Wirtschaft und Technologie
Project number (not EU):03WKR05C
Way of publication:Open Access
Faculty/Chair:Fakultät 2 Umwelt und Naturwissenschaften / FG Zellbiologie
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