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Label-free detection of breast masses using multiphoton microscopy.

Wu X, Chen G, Lu J, Zhu W, Qiu J, Chen J, Xie S, Zhuo S, Yan J - PLoS ONE (2013)

Bottom Line: The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of second-harmonic generation signals.On the contrary, in benign breast masses, second-harmonic generation signals could be seen easily in MPM imaging.These observations indicate that MPM could be an important potential tool to provide label-free noninvasive diagnostic impressions that can guide surgeon in biopsy and patient management.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Fujian Provincial Tumor Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China.

ABSTRACT
Histopathology forms the gold standard for the diagnosis of breast cancer. Multiphoton microscopy (MPM) has been proposed to be a potentially powerful adjunct to current histopathological techniques. A label-free imaging based on two- photon excited fluorescence and second-harmonic generation is developed for differentiating normal breast tissues, benign, as well as breast cancer tissues. Human breast biopsies (including human normal breast tissues, benign as well as breast cancer tissues ) that are first imaged (fresh, unfixed, and unstained) with MPM and are then processed for routine H-E histopathology. Our results suggest that the MPM images, obtained from these unprocessed biopsies, can readily distinguish between benign lesions and breast cancers. In the tissues of breast cancers, MPM showed that the tumor cells displayed marked cellular and nuclear pleomorphism. The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of second-harmonic generation signals. For breast cancer, MPM diagnosis was 100% correct because the tissues of breast cancers did not have second-harmonic generation signals in MPM imaging. On the contrary, in benign breast masses, second-harmonic generation signals could be seen easily in MPM imaging. These observations indicate that MPM could be an important potential tool to provide label-free noninvasive diagnostic impressions that can guide surgeon in biopsy and patient management.

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Schematic illustration of normal breast structure.The main parts of the breast are lobules, ducts, and stroma (fatty tissue and connective tissue surrounding the ducts and lobules). Most breast cancers begin in the ducts (ductal), some in the lobules (lobular) and the rest in other tissues. Fibrocystic breast disease and fibroadenoma usually develop within the breast lobules.
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pone-0065933-g001: Schematic illustration of normal breast structure.The main parts of the breast are lobules, ducts, and stroma (fatty tissue and connective tissue surrounding the ducts and lobules). Most breast cancers begin in the ducts (ductal), some in the lobules (lobular) and the rest in other tissues. Fibrocystic breast disease and fibroadenoma usually develop within the breast lobules.

Mentions: Based on multiphoton images obtained, we were able to differentiate normal breast tissues, benign, as well as breast cancer tissues. Normal breast tissues consisted of the ducts and glands (which maked up the bulk of the volume of the adult breast) as well as adipose and fibrous structures in varying proportions, as shown in Fig. 1. These structures possess strong multiphoton signals and can be clearly recognized in multiphoton images, as shown in Fig. 2A. At high magnification, ducts lined by simple columnar epithelium and obvious basement membrane were identified in the normal tissues with multiphoton microscopy, mainly because of the SHG signals from collagen components, as shown in Fig. 2C. All these features were present in the H-E stained image shown in Fig. 2B, D. Fibrocystic breast changes were characterized by an increase in the number and size of glandular tissues, usually within the breast lobules. These included benign fibrous tissues and scattered cysts containing amorphous material. The cyst lining was flattened or absent in some cases. These features were distinctly observed in the multiphoton image and H-E stained image shown in Fig. 3A, B respectively. It was seen in Figure 4A that fibroadenoma showed unique morphological structure with the acinar arrangement of the round and ovoid tubules, surrounded by a myoepithelial layer and the stromal component. The stroma is made up of loose connective tissue (as shown in Fig. 4C). The same pattern was also clearly identified in the H-E stained image, as shown in Fig. 4B and 4D. Figure.5A showed multiphoton image of breast cancer. The tumor cells displayed marked cellular and nuclear pleomorphism. The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of basement membrane (no SHG signals). Glandular structure consisting of tumor cells was evident, as indicated by the arrow in the multiphoton image shown in Fig. 5C. These features were consistent with that in the H-E stained image shown as Fig. 5D. Comparative features between H-E and MPM were shown in Table 1.


Label-free detection of breast masses using multiphoton microscopy.

Wu X, Chen G, Lu J, Zhu W, Qiu J, Chen J, Xie S, Zhuo S, Yan J - PLoS ONE (2013)

Schematic illustration of normal breast structure.The main parts of the breast are lobules, ducts, and stroma (fatty tissue and connective tissue surrounding the ducts and lobules). Most breast cancers begin in the ducts (ductal), some in the lobules (lobular) and the rest in other tissues. Fibrocystic breast disease and fibroadenoma usually develop within the breast lobules.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3675049&req=5

pone-0065933-g001: Schematic illustration of normal breast structure.The main parts of the breast are lobules, ducts, and stroma (fatty tissue and connective tissue surrounding the ducts and lobules). Most breast cancers begin in the ducts (ductal), some in the lobules (lobular) and the rest in other tissues. Fibrocystic breast disease and fibroadenoma usually develop within the breast lobules.
Mentions: Based on multiphoton images obtained, we were able to differentiate normal breast tissues, benign, as well as breast cancer tissues. Normal breast tissues consisted of the ducts and glands (which maked up the bulk of the volume of the adult breast) as well as adipose and fibrous structures in varying proportions, as shown in Fig. 1. These structures possess strong multiphoton signals and can be clearly recognized in multiphoton images, as shown in Fig. 2A. At high magnification, ducts lined by simple columnar epithelium and obvious basement membrane were identified in the normal tissues with multiphoton microscopy, mainly because of the SHG signals from collagen components, as shown in Fig. 2C. All these features were present in the H-E stained image shown in Fig. 2B, D. Fibrocystic breast changes were characterized by an increase in the number and size of glandular tissues, usually within the breast lobules. These included benign fibrous tissues and scattered cysts containing amorphous material. The cyst lining was flattened or absent in some cases. These features were distinctly observed in the multiphoton image and H-E stained image shown in Fig. 3A, B respectively. It was seen in Figure 4A that fibroadenoma showed unique morphological structure with the acinar arrangement of the round and ovoid tubules, surrounded by a myoepithelial layer and the stromal component. The stroma is made up of loose connective tissue (as shown in Fig. 4C). The same pattern was also clearly identified in the H-E stained image, as shown in Fig. 4B and 4D. Figure.5A showed multiphoton image of breast cancer. The tumor cells displayed marked cellular and nuclear pleomorphism. The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of basement membrane (no SHG signals). Glandular structure consisting of tumor cells was evident, as indicated by the arrow in the multiphoton image shown in Fig. 5C. These features were consistent with that in the H-E stained image shown as Fig. 5D. Comparative features between H-E and MPM were shown in Table 1.

Bottom Line: The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of second-harmonic generation signals.On the contrary, in benign breast masses, second-harmonic generation signals could be seen easily in MPM imaging.These observations indicate that MPM could be an important potential tool to provide label-free noninvasive diagnostic impressions that can guide surgeon in biopsy and patient management.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Fujian Provincial Tumor Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China.

ABSTRACT
Histopathology forms the gold standard for the diagnosis of breast cancer. Multiphoton microscopy (MPM) has been proposed to be a potentially powerful adjunct to current histopathological techniques. A label-free imaging based on two- photon excited fluorescence and second-harmonic generation is developed for differentiating normal breast tissues, benign, as well as breast cancer tissues. Human breast biopsies (including human normal breast tissues, benign as well as breast cancer tissues ) that are first imaged (fresh, unfixed, and unstained) with MPM and are then processed for routine H-E histopathology. Our results suggest that the MPM images, obtained from these unprocessed biopsies, can readily distinguish between benign lesions and breast cancers. In the tissues of breast cancers, MPM showed that the tumor cells displayed marked cellular and nuclear pleomorphism. The tumor cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, infiltrated into disrupted connective tissue, leading to the loss of second-harmonic generation signals. For breast cancer, MPM diagnosis was 100% correct because the tissues of breast cancers did not have second-harmonic generation signals in MPM imaging. On the contrary, in benign breast masses, second-harmonic generation signals could be seen easily in MPM imaging. These observations indicate that MPM could be an important potential tool to provide label-free noninvasive diagnostic impressions that can guide surgeon in biopsy and patient management.

Show MeSH
Related in: MedlinePlus