Ann Dermatol.  2011 Feb;23(1):44-52. 10.5021/ad.2011.23.1.44.

Characterizing Cutaneous Elastic Fibers by Eosin Fluorescence Detected by Fluorescence Microscopy

• Heo YS
• Song HJ

• KMID: 2171951
• DOI: http://doi.org/10.5021/ad.2011.23.1.44

Abstract

BACKGROUND
Though elastic fibers are as important as collagen fibers in interpretation of the histopathologic findings, it is impossible to observe them on the hematoxylin & eosin (H&E) stained specimen.
OBJECTIVE
Characterizing eosin fluorescence emitted by elastic fibers in H&E stained specimens.
METHODS
Normal skin tissue sections were stained in 4 different ways (unstained, hematoxylin only, eosin only, H&E) and observed under a fluorescence microscope using a FITC filter set. Fluorescent findings of 30 H&E-stained specimens showing abnormal dermal findings were compared with bright field findings of Miller's elastic stained specimen.
RESULTS
Strong eosin fluorescence was related to the differential binding property of eosin with elastic fibers. Hematoxylin stain quenched excessive eosin fluorescence from other tissue components and contributed to better contrast. Fluorescence microscopy of H&E-stained sections was found to be especially useful in observing mature elastic fibers in the reticular dermis. In 74% of the specimens, eosin fluorescence findings of elastic fibers in reticular dermis matched well with that of specimens with elastic fiber special stain.
CONCLUSION
Analysis of skin elastic fibers by fluorescence microscopy is a useful and complementary method to reveal hidden elastic fibers in H&E-stained specimens.

Keyword

Elastic fiber; Eosin; Fluorescence

MeSH Terms

Collagen
Dermis
Elastic Tissue
Enzyme Multiplied Immunoassay Technique
Eosine Yellowish-(YS)
Fluorescein-5-isothiocyanate
Fluorescence
Hematoxylin
Microscopy, Fluorescence
Skin
Collagen
Eosine Yellowish-(YS)
Fluorescein-5-isothiocyanate
Hematoxylin

Figure

• 

  Fig. 1 Basic features of the fluorescence of elastic fibers stained with H&E. (A) Elastic fibers are invisible by bright field microscopy (H&E, ×200). (B) Elastic fibers in the same tissue specimen (arrow heads) are visible by fluorescence microscopy (brightly yellow dots and streaks) (H&E, ×200). (C) Miller's elastic stain of elastic fibers shows corresponding pattern in the dermis as observed by fluorescence microscopy (arrow head) (×200).

• 

  Fig. 2 Basic features of eosin fluorescence in different steps of H&E staining process. (A, B) Hematoxylin stain only, (C, D) eosin stain only, (E, F) H&E, (G, H) unstained background, (I) Miller's elastic stain. (D, F) Photographic enlarged view of dotted box area in C, E. (A, C, E, G, I) Bright field microscopic findings (×200), (B, D, F, H) fluorescence microscopic findings (×200).

• 

  Fig. 3 Eosin fluorescence findings of elastic fibers in the dermis. (A, C, E) Elastic fibers (oxytalan and elaunin) in the papillary dermis. (B, D, F) Mature elastic fibers (arrow heads) in the reticular dermis. (A, B) Bright field microscopy (H&E, ×400), (C, D) fluorescence microscopy (H&E, ×400), (E, F) bright field microscopy (Miller's elastic stain, ×400).

• 

  Fig. 4 Loss of fluorescence in photoaged skin. Eosin fluorescence was lost in photo damaged elastic fibers of the papillary dermis. But in Miller's elastic stain, dark clumping degenerated elastic fibers (red arrow) in the papillary dermis stained the same as normal elastic fibers in the reticular dermis. (A) Bright field microscopy (H&E, ×100), (B) fluorescence microscopy (H&E, ×100), (C) bright field microscopy (Miller's elastic stain, ×100).

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  Fig. 5 Eosin fluorescence findings in pseudoxanthoma elasticum. (A) Calcified altered elastic fibers were observed (H&E, ×400). (B) Elastic fibers showed the same pattern as H&E stain (Miller's elastic stain, ×400). (C) Intact elastic fibers (arrow head), degenerative elastic fibers (thick arrow) and brown color calcium depositions (thin arrow) were observed in different pattern to H&E or elastic stain (fluorescence microscopy, ×400).

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