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Fluorescence patterns of autoantibodies

Typical patterns of ANA using HEp-2 cells

Rodent tissue sections which were used as substrates during early periods of ANA testing have been replaced by HEp-2 cells (immortalized cells orginated from human laryngeal carcinoma) grown as a monolayer on the slide. The HEp-2 cells have much larger structures and higher cell division rates, so the detailed nuclear and cytoplasmic staining patterns can be recognized in the cell populations of different cell cycles. It is noted that HEp-2 cells exhibit more than 30 different patterns, some are antigen-specific but some are similar with different specificities, and the mixed patterns by different autoantibodies in patients' sera are frequently observed.


photo 1

No significant nuclear and cytoplasmic staining. Depending on the sensitivity of the fluorescence microscope, faint fluorescence can be observed, which is considered negative.

In case of positive, staining patterns are interpreted.

Homogeneous nuclear staining

photo 2   HEPASERA-1**

Uniform diffuse fluorescence staining of the entire nucleus in interphase cells. The chromosomal regions in mitotic cells give intense fluorescence.

Peripheral nuclear staining

photo 3

Uniform diffuse nuclear staining similar to homogeneous staining, with greater intensity at its outer rim. The chromosomal region in mitotic cells has uniform staining with greater intensity at its outer rim. Most of the sera showing this pattern give homogeneous staining when sera are diluted.

Speckled nuclear staining

photo 4   HEPASERA-1**

Fine or coarse speckles are distributed in the entire nucleus of interphase cells. The chromosomal region shows no flurorescence in most of the mitotic cells.

Centromere (Discrete-speckled) staining

photo 5   HEPASERA-1**

40 to 80 discrete speckles are stained in the nucleus of interphase cells. Speckles aligned in the chromosomal region of mitotic cells can be seen.

Nucleolar staining

photo 6   HEPASERA-1**

Homogeneous or speckled (clumpy) staining of the nucleolus of interphase cells are seen by various nucleolar autoantibodies.

**HEPASERA-1 is MBL's ANA reference sera (Homogeneous pattern, Speckled pattern, Nucleolar pattern, Discrete speckled pattern).

Other staining patterns and antigens,

Cell cycle-dependent
pleomorphic staining
proliferating cell nuclear antigen (PCNA)
Nuclear dots staining minute nuclear antigens; p80 coilin, Sp100 etc.
Nuclear membranous
lamin, nuclear pore complex
Cytoplasmic staining cytoplasmic antigens; enzymes such as aminoacyl-tRNA synthetases, organelles such as mitochondria and ribosome, cytoskelton, mitotic spindle apparatus


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