Label-free detection of breast masses using multiphoton microscopy

PLoS One. 2013 Jun 6;8(6):e65933. doi: 10.1371/journal.pone.0065933. Print 2013.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biopsy
  • Breast Neoplasms / diagnosis*
  • Female
  • Humans
  • In Vitro Techniques
  • Microscopy, Fluorescence, Multiphoton / methods*

Grants and funding

This work was supported by the National Natural Science Foundation of China (81272574, 61275006 and 81271620), the Program for Changjiang Scholars and Innovative Research Team in University (Grant No. IRT1115), the Natural Science Foundation of Fujian Province (2010J01136 and 2011J01341), and the Program from Education Bureau of Fujian Province (JA12057). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.