Sentinel lymph node (SLN) mapping is a valuable and crucial diagnostic procedure in staging malignancies. We compared two non‐invasive techniques, near‐infrared (NIR) fluorescence imaging and contrast‐enhanced ultrasound (CEUS), to identify the SLNs in three superficial anatomical regions in an animal model. Six healthy laboratory dogs were included in a proof‐of‐concept trial. A NIR fluorescent dye (Indocyanine Green) and microbubbles (Sonovue) were consecutively injected subdermally in the Inguinal, axillary and popliteal region to map the SLNs. Transcutaneous NIR fluorescence imaging identified SLNs in 17 out of a total of 18 occasions. CEUS identified SLNs in all regions (18/18). Whereas NIR fluorescence imaging performed better in the visualization of the afferent lymphatic tract, CEUS demonstrated different filling patterns of the SLNs, a feature potentially critical for the concept of SLN mapping in cancer patients. Both NIR fluorescence imaging and CEUS are safe, non‐invasive, practical and accurate methods to perform real‐time transcutaneous SLN mapping with potential in a clinical setting.

One of the major challenges in surgical oncology is the intraoperative discrimination of tumoural versus healthy tissue. Until today, surgeons rely on visual inspection and palpation to define the tumoural margins during surgery and, unfortunately, for various cancer types, the local recurrence rate thus remains unacceptably high. Near-infrared (NIR) fluorescence imaging is an optical imaging technique that can provide real-time preoperative and intraoperative information after administration of a fluorescent probe that emits NIR light once exposed to a NIR light source. This technique is safe, cost-effective and technically easy. Several NIR fluorescent probes are currently studied for their ability to highlight neoplastic cells. In addition, NIR fluorescence imaging holds great promise for sentinel lymph node mapping. The aim of this manuscript is to provide a literature review of the current organic NIR fluorescent probes tested in the light of human oncology and to introduce fluorescence imaging as a valuable asset in veterinary oncology.

BackgroundNear-infrared fluorescence (NIRF) imaging is a relatively novel technique that can aid surgeons during intraoperative tumour identification.MethodsNine canine oncology patients (five mammary gland tumours, three mast cell tumours and one melanoma) received intravenous indocyanine green (ICG). After 24 hours, tumours were resected and fluorescence intensities of tumours and surroundings were evaluated. Additional wound bed tissue was resected if residual fluorescence was present after tumour resection. Ex vivo, fluorescence-guided dissection was performed to separate tumour from surrounding tissue.ResultsIntraoperative NIRF-guided tumour delineation was feasible in four out of nine dogs. Wound bed imaging after tumour removal identified nine additional fluorescent lesions, of which four contained tumour tissue. One of these four true positive in vivo lesions was missed by standard-of-care inspection. Ex vivo fluorescence-guided tumour dissection showed a sensitivity of 72 per cent and a specificity of 80 per cent in discriminating between tumour and surrounding tissue.ConclusionThe value of ICG for intraoperative tumour delineation seems more limited than originally thought. Although NIRF imaging using ICG did identify remaining tumour tissue in the wound bed, a high false positive rate was also observed.

Shear wave elastography (SWE) is a novel ultrasound imaging technique, used in human medicine to differentiate metastatic from non‐metastatic lymph nodes (LNs) based on higher tissue stiffness. The objective of this pilot study was to evaluate whether canine metastatic LNs were stiffer compared to non‐metastatic LNs. SWE of sentinel or regional LNs in dogs with head and neck cancer was performed. Ten elastograms of each LN were acquired. In each elastogram, mean, median, and maximum shear wave velocities (SWVs) were calculated in the most elastic region, the stiffest region, and the entire LN. The means of those SWVs for each region of each LN were subsequently calculated. Furthermore, a stepwise subsampling was performed to assess the effect of the number of acquired elastograms on the consistency of the classification of a LN as metastatic or not. Twenty‐four LNs in 15 dogs with head and neck cancer were included, of which 10 LNs were metastatic. Metastatic medial retropharyngeal and mandibular LNs were successfully distinguished from non‐metastatic LNs based on higher mean and median SWVs in the stiffest region and higher maximum SWVs in entire LNs. Furthermore, maximum SWVs in the stiffest region of mandibular LNs were higher in metastatic LNs compared to non‐metastatic LNs. Downstream analyses demonstrated that at least eight elastograms were necessary to perform reliable analyses. This pilot trial demonstrated that SWE has potential to discriminate metastatic from non‐metastatic LNs; however, LN status should be based on at least eight elastograms in future clinical trials.