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Two-Photon Microscopy of Stroma <i>ex vivo</i> for the Express Diagnostics of Breast Cancer

Two-Photon Microscopy of Stroma ex vivo for the Express Diagnostics of Breast Cancer

Sergeeva E.A., Kirillin M.Yu., Dudenkova V.V., Pavlov M.V., Orlinskaya N.Yu., Shakhova N.M.
Key words: two-photon microscopy; second harmonic generation; two-photon excited autofluorescence; breast; stroma; fibroadenoma; carcinoma.
2019, volume 11, issue 3, page 89.

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The goal of the study is to develop the approach for the express analysis of ex vivo biopsy specimens of breast tissue based on two-photon microscopy (TPM).

Materials and Methods. We studied breast tissue samples obtained by trephine biopsy procedure. TPM images of breast stroma obtained from the ex vivo biopsy specimens were compared with TPM images of the unstained deparaffinized 10-μm thick histological sections of breast tissue. The study included samples of patients with fibroadenoma (2 cases), in situ carcinoma (1 case), and invasive carcinoma of non-specific type I–II grade (4 cases). In the frames of numerical processing of TPM images, collagen disorganization factor maps based on spatial Fourier analysis were constructed.

Results. We have demonstrated the feasibility of the TPM technique in detecting stroma remodeling in breast biopsy specimens in the development of benign and malignant changes. Typical images of biopsy specimens and histological sections were obtained for normal breast tissue, fibroadenoma and various types of carcinomas. Distinction between typical images of benign and malignant tumors was demonstrated. Changes in collagen fibers structure and the presence/absence of elastin fibers in the considered pathological cases were revealed. Differences in TPM images of biopsy specimens and histological sections are shown, which may originate from the distortion of fiber shape in the course of histological sample preparation. The proposed algorithm for quantitative TPM image processing allows objectifying the results of imaging, which is an essential step towards automated express diagnostics.

Conclusion. Two-photon microscopy has high potential as the express biopsy technique for the primary diagnosis of breast tissue, as well as for intraoperative use.

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