1：Experimental Design and Method Selection:

A first-in-human pilot study was conducted using optical specimen mapping (OSM) with a closed-field fluorescence imaging device. The study involved infusing patients with panitumumab-IRDye800, a fluorescently labeled antibody, to target tumor tissue. The OSM device was used to image resected specimens, and results were correlated with pathological findings.

2：Sample Selection and Data Sources:

Eight patients with head and neck squamous cell carcinoma (HNSCC) were enrolled. Specimens were obtained from surgical resections, and data points (n=160) were extracted from histological sections.

3：List of Experimental Equipment and Materials:

OSM device (IGP-ELVIS, LI-COR), panitumumab-IRDye800, tissue-mimicking phantoms made with agarose, bovine blood, intralipid emulsion, formalin for fixation, hematoxylin and eosin (H&E) staining materials, ImageJ software, Image Studio software (LI-COR Biosciences), LabVIEW control program.

4：Experimental Procedures and Operational Workflow:

Patients were infused with panitumumab-IRDye800 1-4 days before surgery. After resection, specimens were imaged with the OSM device in the operating room. Specimens were then formalin-fixed, sectioned into 5 mm slices, processed, and H&E stained. Fluorescence signals were analyzed using thresholding, and margin distances were measured from digitized H&E slides.

5：Data Analysis Methods:

Quantitative analysis of fluorescence intensity was performed using Image Studio. Sensitivity and specificity were calculated using receiver operating characteristic (ROC) curves. Statistical analysis included unpaired t-tests for comparing margin distances, with results reported as means with standard deviations and 95% confidence intervals.