Abstract
BackgroundIn over 20% of breast conserving operations, postoperative pathological assessment of the excised tissue reveals positive margins, requiring additional surgery. Current techniques for intra-operative assessment of tumor margins are insufficient in accuracy or resolution to reliably detect small tumors. There is a distinct need for a fast technique to accurately identify tumors smaller than 1 mm2 in large tissue surfaces within 30 min.MethodsMulti-modal spectral histopathology (MSH), a multimodal imaging technique combining tissue auto-fluorescence and Raman spectroscopy was used to detect microscopic residual tumor at the surface of the excised breast tissue. New algorithms were developed to optimally utilize auto-fluorescence images to guide Raman measurements and achieve the required detection accuracy over large tissue surfaces (up to 4 × 6.5 cm2). Algorithms were trained on 91 breast tissue samples from 65 patients.ResultsIndependent tests on 121 samples from 107 patients - including 51 fresh, whole excision specimens - detected breast carcinoma on the tissue surface with 95% sensitivity and 82% specificity. One surface of each uncut excision specimen was measured in 12–24 min. The combination of high spatial-resolution auto-fluorescence with specific diagnosis by Raman spectroscopy allows reliable detection even for invasive carcinoma or ductal carcinoma in situ smaller than 1 mm2.ConclusionsThis study provides evidence that this multimodal approach could provide an objective tool for intra-operative assessment of breast conserving surgery margins, reducing the risk for unnecessary second operations.
Highlights
We have demonstrated the feasibility of multimodal spectral histopathology (MSH), a selective-sampling technique that combines high-resolution wide-field auto-fluorescence (AF) microscopy and Raman spectroscopy to detect ductal carcinomas in frozen breast micro-sections (5 × 5 mm2) [38]
Through independent tests of these algorithms and measurement protocols on breast tissue samples and real, whole Breast conserving surgery (BCS) specimen surfaces, we show that MSH has great potential for objective, intra-operative assessment of the excision surface of BCS specimens immediately after excision without requiring any sample preparation
The samples were submitted for histological processing and hematoxylin and eosin (H&E) sections were obtained for each measured surface
Summary
In over 20% of breast conserving operations, postoperative pathological assessment of the excised tissue reveals positive margins, requiring additional surgery. The goal of BCS is to remove the entire tumor while leaving healthy breast tissue intact, providing better cosmetic outcome. This is challenging because of the lack of tools available for intra-operative assessment of margins to indicate complete tumor excision. In more than 20% of BCS procedures, positive margins are detected (i.e. tumor close to the edge) and additional operations are required to achieve complete excision [2, 3] Half of these “re-excisions” are for “on-ink” margins [3], meaning that tumor was found on the surface of the excised tissue. Guidelines from the Society of Surgical Oncology and the American Society for Radiation Oncology state that clear on-ink margins are sufficient to remove tumor and more widely clear margins did not significantly increase the risk of recurrence [4, 5]
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