Hyperspectral dark-field microscopy of human breast lumpectomy samples for tumor margin detection in breast-conserving surgery.

Significance: Hyperspectral dark-field microscopy (HSDFM) and data cube analysis algorithms demonstrate successful detection and classification of various tissue types, including carcinoma regions in human post-lumpectomy breast tissues excised during breast-conserving surgeries. Aim: We expand the application of HSDFM to the classification of tissue types and tumor subtypes in pre-histopathology human breast lumpectomy samples. Approach: Breast tissues excised during breast-conserving surgeries were imaged by the HSDFM and analyzed. The performance of the HSDFM is evaluated by comparing the backscattering intensity spectra of polystyrene microbead solutions with the Monte Carlo simulation of the experimental data. For classification algorithms, two analysis approaches, a supervised technique based on the spectral angle mapper (SAM) algorithm and an unsupervised technique based on the K-means algorithm are applied to classify various tissue types including carcinoma subtypes. In the supervised technique, the SAM algorithm with manually extracted endmembers guided by H&E annotations is used as reference spectra, allowing for segmentation maps with classified tissue types including carcinoma subtypes. Results: The manually extracted endmembers of known tissue types and their corresponding threshold spectral correlation angles for classification make a good reference library that validates endmembers computed by the unsupervised K-means algorithm. The unsupervised K-means algorithm, with no a priori information, produces abundance maps with dominant endmembers of various tissue types, including carcinoma subtypes of invasive ductal carcinoma and invasive mucinous carcinoma. The two carcinomas' unique endmembers produced by the two methods agree with each other within <2% residual error margin. Conclusions: Our report demonstrates a robust procedure for the validation of an unsupervised algorithm with the essential set of parameters based on the ground truth, histopathological information. We have demonstrated that a trained library of the histopathology-guided endmembers and associated threshold spectral correlation angles computed against well-defined reference data cubes serve such parameters. Two classification algorithms, supervised and unsupervised algorithms, are employed to identify regions with carcinoma subtypes of invasive ductal carcinoma and invasive mucinous carcinoma present in the tissues. The two carcinomas' unique endmembers used by the two methods agree to <2% residual error margin. This library of high quality and collected under an environment with no ambient background may be instrumental to develop or validate more advanced unsupervised data cube analysis algorithms, such as effective neural networks for efficient subtype classification.

Micro-computed tomography enables rapid surgical margin assessment during breast conserving surgery (BCS): correlation of whole BCS micro-CT readings to final histopathology.

BACKGROUND: Roughly 23% of breast conserving surgery (BCS) patients undergo a second re-excision procedure due to pathologically positive surgical margins. We investigated the feasibility and potential value of micro-Computed Tomography (micro-CT) as a surgical margin guidance tool during BCS. METHODS: A cohort of 32 BCS specimens was prospectively imaged with a pre-clinical micro-CT system upon arrival in the surgical pathology laboratory. Reconstructed micro-CT scans were evaluated retrospectively by an experienced breast radiologist, who provided binary determinations whether lesions extended to the specimen margin. These readings were then compared to the final pathological diagnosis and to 2D specimen radiography readings. RESULTS: Of the 32 specimens imaged, 28 had malignant and four had benign pathological diagnoses. Overall five (four malignant, one benign) of the 32 specimens had lesion tissue extending to the margin. For all 32 specimens, micro-CT reconstructions were calculated (< 4 min. acquisition + reconstruction time) and each specimen was volumetrically analyzed by a radiologist. Of the 28 malignant specimen readings, 18 matched the final pathological diagnosis [64%, 95 CI (47%-81%)], with a negative predictive value of 89% [95 CI (74%-96%)]. Micro-CT readings revealed changes in the tumor location and margin status as compared to single-projection radiography readings. CONCLUSIONS: Micro-CT scanning of BCS specimens enabled margin status assessment over the entirety of the surgical surface in a clinically relevant time frame, provided additional spatial information over single-projection radiography, and may be a potentially useful BCS guidance tool.

Molecular dyes used for surgical specimen margin orientation allow for intraoperative optical assessment during breast conserving surgery.

A variety of optical techniques utilizing near-infrared (NIR) light are being proposed for intraoperative breast tumor margin assessment. However, immediately following a lumpectomy excision, the margins are inked, which preserves the orientation of the specimen but prevents optical interrogation of the tissue margins. Here, a workflow is proposed that allows for both NIR optical assessment following full specimen marking using molecular dyes which have negligible absorption and scattering in the NIR. The effect of standard surgical inks in contrast to molecular dyes for an NIR signal is shown. Further, the proposed workflow is demonstrated with full specimen intraoperative imaging on all margins directly after the lumpectomy has been excised and completely marked. This work is an important step in the path to clinical feasibility of intraoperative breast tumor margin assessment using NIR optical methods without having to compromise on the current clinical practice of inking resected specimens for margin orientation.

Projection imaging of photon beams by the Cerenkov effect.

PURPOSE: A novel technique for beam profiling of megavoltage photon beams was investigated for the first time by capturing images of the induced Cerenkov emission in water, as a potential surrogate for the imparted dose in irradiated media. METHODS: A high-sensitivity, intensified CCD camera (ICCD) was configured to acquire 2D projection images of Cerenkov emission from a 4 × 4 cm(2) 6 MV linear accelerator (LINAC) x-ray photon beam operating at a dose rate of 400 MU∕min incident on a water tank with transparent walls. The ICCD acquisition was gated to the LINAC sync pulse to reduce background light artifacts, and the measurement quality was investigated by evaluating the signal to noise ratio and measurement repeatability as a function of delivered dose. Monte Carlo simulations were used to derive a calibration factor for differences between the optical images and deposited dose arising from the anisotropic angular dependence of Cerenkov emission. Finally, Cerenkov-based beam profiles were compared to a percent depth dose (PDD) and lateral dose profile at a depth of d(max) from a reference dose distribution generated from the clinical Varian ECLIPSE treatment planning system (TPS). RESULTS: The signal to noise ratio was found to be 20 at a delivered dose of 66.6 cGy, and proportional to the square root of the delivered dose as expected from Poisson photon counting statistics. A 2.1% mean standard deviation and 5.6% maximum variation in successive measurements were observed, and the Monte Carlo derived calibration factor resulted in Cerenkov emission images which were directly correlated to deposited dose, with some spatial issues. The dose difference between the TPS and PDD predicted by Cerenkov measurements was within 20% in the buildup region with a distance to agreement (DTA) of 1.5-2 mm and ±3% at depths beyond d(max). In the lateral profile, the dose difference at the beam penumbra was within ±13% with a DTA of 0-2 mm, ±5% in the central beam region, and 2%-3% in the beam umbra. CONCLUSIONS: The results from this initial study demonstrate the first documented use of Cerenkov emission imaging to profile x-ray photon LINAC beams in water. The proposed modality has several potential advantages over alternative methods, and upon future refinement may prove to be a robust and novel dosimetry method.