Establishment of HEp-2 cell preparation for automated analysis of ANA fluorescence pattern.
BACKGROUND
Identification 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.


METHODS
HEp-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.


RESULTS
The 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.


CONCLUSIONS
After 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.
Hahm
Daniel
D
Cell biology and Tissue Engineering, Lausitz University of Applied Sciences, Senftenberg, Germany.
Anderer
Ursula
U
D000974:Antibodies, Antinuclear / Q000032:analysis*; D045744:Cell Line, Tumor; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D005404:Fixatives / Q000737:chemistry; D006416:Hematoxylin / Q000737:chemistry; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D007211:Indoles / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods; D013194:Staining and Labeling / Q000379:methods
D000974:Antibodies, Antinuclear; D005404:Fixatives; D007211:Indoles; C007293:DAPI; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin