Original Article

Subject Category: Melanocytes/Melanoma

Journal of Investigative Dermatology (2008) 128, 1248–1255; doi:10.1038/sj.jid.5701139; published online 8 November 2007

Multiphoton Laser Scanning Microscopy on Non-Melanoma Skin Cancer: Morphologic Features for Future Non-Invasive Diagnostics

John Paoli1, Maria Smedh2, Ann-Marie Wennberg1 and Marica B Ericson1,3

  1. 1Department of Dermatology, Sahlgrenska University Hospital, Göteborg University, Göteborg, Sweden
  2. 2Centre for Cellular Imaging, Göteborg University, Göteborg, Sweden
  3. 3Department of Physics, Göteborg University, Göteborg, Sweden

Correspondence: Dr John Paoli, Department of Dermatology, Sahlgrenska University Hospital, Göteborg 413 45, Sweden. E-mail: john.paoli@vgregion.se

Received 8 March 2007; Accepted 16 August 2007; Published online 8 November 2007.

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Abstract

This study describes the morphologic features of human non-melanoma skin cancer obtained using multiphoton laser scanning microscopy (MPLSM) on freshly excised specimens from 14 patients. Optical sectioning parallel to the tissue surface was performed, resulting in en face autofluorescence images of the epidermis and upper dermis, reaching tissue depths of 135 mum. The microscopy was carried out ex vivo using a femtosecond pulsed laser at 780 nm and a times 40/0.8 objective. The autofluorescence was detected in the range of 450–530 nm. Traditional histopathological criteria such as bowenoid dysplasia, multinucleated cells, or hyperkeratosis in squamous cell carcinoma in situ (SCCIS) (five specimens), and peripheral palisading of tumor cells in superficial basal cell carcinoma (SBCC) (six specimens) were clearly discerned. The morphologic features differed significantly between these lesions and perilesional skin. However, characteristic tumor aggregates were found in only one of the three investigated nodular basal cell carcinomas (NBCCs) due to limited imaging depth. In addition, speckled perinuclear fluorescence was observed in both lesions and normal perilesional skin. In conclusion, MPLSM could potentially be applied for non-invasive diagnostics of SCCIS and SBCC, whereas the ability to characterize NBCC is unclear at this point.

Abbreviations:

BCC, basal cell carcinoma; DP, dermal papillae; MPLSM, multiphoton laser scanning microscopy; NADPH, nicotinamide adenine dinucleotide phosphate; NBCC, nodular basal cell carcinoma; NIR, near-infrared; RCLSM, reflectance-mode confocal laser scanning microscopy; SB, stratum basale; SBCC, superficial basal cell carcinoma; SC, stratum corneum; SCCIS, squamous cell carcinoma in situ; SG, stratum granulosum; SS, stratum spinosum

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