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. 2008 Jan-Feb;10(1):38-43.
doi: 10.1001/archfacial.2007.18.

Nondestructive imaging of live human keloid and facial tissue using multiphoton microscopy

Victor Da Costa  1 , Randy Wei, Ryan Lim, Chung-Ho Sun, Jimmy J Brown, Brian J-F Wong
Affiliations

Nondestructive imaging of live human keloid and facial tissue using multiphoton microscopy

Victor Da Costa et al. Arch Facial Plast Surg. 2008 Jan-Feb.

Abstract

Objectives: To use multiphoton microscopy to image collagen fibers and matrix structure in nonfixed human keloid tissue and normal human facial skin obtained following surgery and to compare the findings to existing knowledge of normal skin and keloid morphology to determine if this technology is a suitable adjunct for conventional histology.

Methods: Epidermis was removed to expose the fibroblast-rich dermal layer that was then imaged using a multiphoton confocal microscope (Zeiss-Meta 510; Carl Zeiss, Jena, Germany). An 800-nm tunable titanium/sapphire femtosecond laser (Mai-Tai; Newport Co Spectra-Physics, Mountain View, California) was used to excite the tissue; second harmonic generation between 397 and 408 nm and autofluorescent signals were collected. Images were obtained using a Plan-Neofluar x40 oil immersion objective lens and a Plan-Apochromat x63 oil immersion lens.

Results: Compared with normal skin, keloids showed disorganized collagen fibers arranged in complex swirls and bundles 20 to 30 microm in diameter. Normal tissue showed collagen fibers as distinct, straight strands less than 10 microm in diameter. Differences between normal and keloid tissue were subtle but apparent.

Conclusions: The value of imaging living tissue is a significant benefit. Because keloids and hypertrophic scars result from altered collagen metabolism, the development of clinical multiphoton microscopy systems may allow examination of wound healing dynamics in vivo and potentially provides a means to monitor therapy without the need for biopsy or the risk of injury to tissue.

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Figures

Figure 1
Figure 1
Serial z-stack images of keloid tissue at a ×63 oil immersion objective lens. Sections are 2.1 µm each and extend 31.2 µm into the surface of the tissue.
Figure 2
Figure 2
Multiphoton confocal microscopic images from normal human facial skin (A and B) and human keloid tissue (C and D) shown under a ×40 oil immersion objective lens (A and C) and a ×63 oil immersion objective lens (B and D).
Figure 3
Figure 3
Multiphoton confocal microscopic images at a ×63 oil immersion objective lens for human keloid tissue (A) and normal human facial skin (B).
See this image and copyright information in PMC

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