Human tissues were obtained under a protocol approved by the Institutional Review Boards at the University of Illinois at Urbana-Champaign and Carle Foundation Hospital, Urbana, Illinois. A total of 29 breast cancer human subjects and 7 healthy (and no history of cancer) human subjects undergoing breast reduction surgeries were included in this study (Table 1). During the breast cancer surgeries, and immediately following resection, the fresh human breast tissue specimens were directly passed to the research team member in the operating room for ex vivo imaging. On the basis of guidance from the surgeon, the location on the surgical margin surface that was deemed to be closest to the tumor within the resected mass was identified and selected as the site from which images were collected. The distance between the imaged surgical margin surface and the tumor mass (tumor-to-margin distance) was later measured postoperatively by the pathologists.

A total of 29 breast cancer subjects and 7 healthy cancer-free subjects undergoing breast reduction surgery were included. Healthy subjects are indicated by NML (normal) under the histologic grade column.

The multimodal label-free images of the unperturbed human breast specimens were acquired within a time window of less than 30 min between the time of surgical excision and tissue fixation for histopathological processing, without disrupting or delaying the surgical procedure. Following imaging, the imaged regions on the surgical margin surfaces of the breast tissue specimens were marked with surgical ink for later registration with histological slides for image feature correlations. After intraoperative imaging, the intact specimens were sent to the pathology laboratory for standard processing and diagnosis.

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