5 years ago

Phasor–FLIM as a Screening Tool for the Differential Diagnosis of Actinic Keratosis, Bowen’s Disease, and Basal Cell Carcinoma

Phasor–FLIM as a Screening Tool for the Differential Diagnosis of Actinic Keratosis, Bowen’s Disease, and Basal Cell Carcinoma
Yuan Lu, Teng Luo, Shaoxiong Liu, Junle Qu, Danying Lin
The aim of this study was to distinguish basal cell carcinoma (BCC) from actinic keratosis (AK) and Bowen’s disease (BD) by fluorescence lifetimes of hematoxylin and eosin (H&E) and phasor analysis. Pseudocolor images of average fluorescence lifetime (τm) exhibited more contrast than conventional bright field and/or fluorescence images of H&E-stained sections. The mean values (μ) of τm distribution (τmμ) in three layers of skin were first explored for comparison with the corresponding layers of AK, BD, and BCC. Moreover, analysis of the H&E fluorescence lifetimes in the phasor space was performed by observing clusters in specific regions of the phasor plot. Various structures in the skin were distinguished. Comparisons of phase distributions from the corresponding layers of skin resulted in quantitative separation and calculation of distinctive parameters including coordinate values, diagonal slopes, and phasor areas. The combination of fluorescence lifetime imaging microscopy (FLIM) and phasor approach (phasor–FLIM) provides a simple method for histopathology analysis and can significantly improve the accuracy of bright field H&E diagnosis. We therefore believe that phasor–FLIM is an aided tool with the potential to provide rapid confirmation of diagnostic criteria and classification of histological types of skin neoplasms.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b01681

DOI: 10.1021/acs.analchem.7b01681

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