Compact portable multiphoton microscopy reveals histopathological hallmarks of unprocessed lung tumor tissue in real time

During lung cancer operations a rapid and reliable assessment of tumor tissue can reduce operation time and potentially improve patient outcomes. We show that third harmonic generation (THG), second harmonic generation (SHG) and two‐photon excited autofluorescence (2PEF) microscopy reveals relevant, histopathological information within seconds in fresh unprocessed human lung samples. We used a compact, portable microscope and recorded images within 1 to 3 seconds using a power of 5 mW. The generated THG/SHG/2PEF images of tumorous and nontumorous tissues are compared with the corresponding standard histology images, to identify alveolar structures and histopathological hallmarks. Cellular structures (tumor cells, macrophages and lymphocytes) (THG), collagen (SHG) and elastin (2PEF) are differentiated and allowed for rapid identification of carcinoid with solid growth pattern, minimally enlarged monomorphic cell nuclei with salt‐and‐pepper chromatin pattern, and adenocarcinoma with lipidic and micropapillary growth patterns. THG/SHG/2PEF imaging is thus a promising tool for clinical intraoperative assessment of lung tumor tissue

Plasticity between Epithelial and Mesenchymal States Unlinks EMT from Metastasis-Enhancing Stem Cell Capacity

Forced overexpression and/or downregulation of proteins regulating epithelial-to-mesenchymal transition (EMT) has been reported to alter metastasis by changing migration and stem cell capacity of tumor cells. However, these manipulations artificially keep cells in fixed states, while in vivo cells may adapt transient and reversible states. Here, we have tested the existence and role of epithelial-mesenchymal plasticity in metastasis of mammary tumors without artificially modifying EMT regulators. In these tumors, we found by intravital microscopy that the motile tumor cells have undergone EMT, while their epithelial counterparts were not migratory. Moreover, we found that epithelial-mesenchymal plasticity renders any EMT-induced stemness differences, as reported previously, irrelevant for metastatic outgrowth, because mesenchymal cells that arrive at secondary sites convert to the epithelial state within one or two divisions, thereby obtaining the same stem cell potential as their arrived epithelial counterparts. We conclude that epithelial-mesenchymal plasticity supports migration but additionally eliminates stemness-enhanced metastatic outgrowth differences

Bringing third and second harmonic generation microscopy into the clinic for the assessment of fresh lung tissue

Third and second harmonic generation (THG/SHG) microscopy is a novel imaging technique that has been suggested as a promising clinical tool, mainly for cancer diagnosis. Here, we test for the first time a compact, transportable ex-vivo THG/SHG microscope (Tritos Diagnostics B.V.) for clinical use. In lung surgery, immediate feedback on the nature of the excised tissue is important. We will present the first results of an ongoing study in which we image both lung tumor tissue immediately after excision and frozen section biopsies, and compare the images to the golden standard histology. Prior to this study, we performed a set of experiments collecting 3D images of mouse lung tissue to show that the combination of THG and SHG microscopy reveals key lung morphology components

Bringing third and second harmonic generation microscopy into the clinic for the assessment of fresh lung tissue

Third and second harmonic generation (THG/SHG) microscopy is a novel imaging technique that has been suggested as a promising clinical tool, mainly for cancer diagnosis. Here, we test for the first time a compact, transportable ex-vivo THG/SHG microscope (Tritos Diagnostics B.V.) for clinical use. In lung surgery, immediate feedback on the nature of the excised tissue is important. We will present the first results of an ongoing study in which we image both lung tumor tissue immediately after excision and frozen section biopsies, and compare the images to the golden standard histology. Prior to this study, we performed a set of experiments collecting 3D images of mouse lung tissue to show that the combination of THG and SHG microscopy reveals key lung morphology components

Bringing third and second harmonic generation microscopy into the clinic for the assessment of fresh lung tissue

Third and second harmonic generation (THG/SHG) microscopy is a novel imaging technique that has been suggested as a promising clinical tool, mainly for cancer diagnosis. Here, we test for the first time a compact, transportable ex-vivo THG/SHG microscope (Tritos Diagnostics B.V.) for clinical use. In lung surgery, immediate feedback on the nature of the excised tissue is important. We will present the first results of an ongoing study in which we image both lung tumor tissue immediately after excision and frozen section biopsies, and compare the images to the golden standard histology. Prior to this study, we performed a set of experiments collecting 3D images of mouse lung tissue to show that the combination of THG and SHG microscopy reveals key lung morphology components