Abstract
Non-melanoma skin cancer (NMSC) belongs to the most frequent human neoplasms. Its exposed location facilitates a fast ambulant treatment. However, in the clinical practice far more lesions are removed than necessary, due to the lack of an efficient pre-operational examination procedure: Standard imaging methods often do not provide a sufficient spatial resolution. The demand for an efficient in vivo imaging technique might be met in the near future by non-linear microscopy. As a first step towards this goal, the appearance of NMSC in various microspectroscopic modalities has to be defined and approaches have to be derived to distinguish healthy skin from NMSC using non-linear optical microscopy. Therefore, in this contribution the appearance of ex vivo NMSC in a combination of coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG) and two photon excited fluorescence (TPEF) imaging—referred as multimodal imaging—is described. Analogous to H&E staining, an overview of the distinct appearances and features of basal cell and squamous cell carcinoma in the complementary modalities is derived, and is expected to boost in vivo studies of this promising technological approach.
Highlights
At present non-melanoma skin cancer (NMSC) accounts for about 20% of all newly detected malignant neoplasms in humans [1]
The joined laser beams are coupled into an inverse laser scanning microscope (LSM 510 Meta, Zeiss, Jena, Germany) and focused at the sample with a 20×/NA 0.8 achromatic objective (Zeiss).The signal detection is performed using photomultipliers (Hamamatsu Photonics, Hamamatsu, Japan) in forward direction (CARS, second harmonic generation (SHG)) and backward direction (TPEF).Both single pictures and large area scans were recorded: The large area scans are composed of up to 15 × 15 tiles, each having a size of 450 μm × 450 μm with a resolution of 1,024 × 1,024 pixels
To guarantee perfect co-registration of the images all three distinct signals, i.e., coherent anti-Stokes Raman scattering (CARS), two photon excited fluorescence (TPEF) and SHG could be detected simultaneous in future measurements
Summary
At present non-melanoma skin cancer (NMSC) accounts for about 20% of all newly detected malignant neoplasms in humans [1]. At the beginning of the 21st century, the number of annual incidences of NMSC varied between 50 and 2,000 among 100,000 individuals in Finland and Australia [2]. The mortality rate for non-melanoma skin cancer remains low compared to other types of cancer [4]. This can be ascribed to a low metastatic potential, and the efficient and effective ambulant treatment, which is possible due to the exposed position of skin cancer. The precondition for the appropriate treatment, remains the detection of the skin neoplasm and its accurate pre-surgery classification. The sensitivity for clinical diagnosis of NMSC is estimated to be about 56% to 90% while the specificity is in the range of 75% to
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