Second and third harmonic generation microscopy visualizes key structural components in fresh unprocessed healthy human breast tissue

Real-time assessment of excised tissue may help to improve surgical results in breast tumor surgeries. Here, as a step towards this purpose, the potential of second and third harmonic generation (SHG, THG) microscopy is explored. SHG and THG are nonlinear optical microscopic techniques that do not r...

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Bibliographic Details
Published in:Journal of biophotonics 2019-06, Vol.12 (6), p.e201800297-e201800297
Main Authors: van Huizen, Laura M G, Kuzmin, Nikolay V, Barbé, Ellis, van der Velde, Susanne, Te Velde, Elisabeth A, Groot, Marie Louise
Format: Article
Language:English
Subjects:
Animals
Blood vessels
Breast
Breast - cytology
Breast - diagnostic imaging
Breast cancer
Connective tissues
Deoxyribonucleic acid
DNA
Ducts
Female
Fluorescence
Full
Harmonic generations
Histology
Histopathology
Human tissues
Humans
Image Processing, Computer-Assisted
Image quality
Mice
Microscopy
Nuclei (cytology)
Second Harmonic Generation Microscopy - methods
Sectioning
Stroma
Surgery
Surgical instruments
Tissues
Tumors
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Summary:Real-time assessment of excised tissue may help to improve surgical results in breast tumor surgeries. Here, as a step towards this purpose, the potential of second and third harmonic generation (SHG, THG) microscopy is explored. SHG and THG are nonlinear optical microscopic techniques that do not require labeling of tissue to generate 3D images with intrinsic depth-sectioning at sub-cellular resolution. Until now, this technique had been applied on fixated breast tissue or to visualize the stroma only, whereas most tumors start in the lobules and ducts. Here, SHG/THG images of freshly excised unprocessed healthy human tissue are shown to reveal key breast components-lobules, ducts, fat tissue, connective tissue and blood vessels, in good agreement with hematoxylin and eosin histology. DNA staining of fresh unprocessed mouse breast tissue was performed to aid in the identification of cell nuclei in label-free THG images. Furthermore, 2- and 3-photon excited auto-fluorescence images of mouse and human tissue are collected for comparison. The SHG/THG imaging modalities generate high quality images of freshly excised tissue in less than a minute with an information content comparable to that of the gold standard, histopathology. Therefore, SHG/THG microscopy is a promising tool for real-time assessment of excised tissue during surgery.
ISSN:1864-063X
1864-0648
DOI:10.1002/jbio.201800297