This proof-of-concept study demonstrates that higher harmonic generation (HHG) microscopy enables rapid, label-free imaging of transbronchial lung cryobiopsies in suspected ILD. On-site within minutes, HHG revealed alveolar structures, fibrosis, and inflammation. This allowed pathologists to assess sample adequacy, and findings were in good concordance with subsequent H&E histology. The results highlight HHG’s potential to reduce sampling errors and speed up diagnostic workflows.
Assessing sample adequacy in ILD
This study shows that a portable HHG microscope, combined with an AI-analysis algorithm, can deliver on-site histologic feedback on lung and pleural biopsies in about 6 minutes. Among 109 biopsies from 47 patients, 97% had sufficient image quality for diagnosing malignancy or non-malignancy, and pathologists correctly classified ~87% of images. This work supports HHG + AI as a fast, reliable tool to accelerate diagnosis and guide treatment decisions.
On-site histological feedback on lung and pleural biopsies
Lack of a clear tumor boundary hampers the surgeon’s ability to accurately detect and resect infiltrating tumor tissue. Higher harmonic generation enables label-free imaging of excised brain tissue, revealing histological hallmarks within seconds. The combination of real-time imaging and image analysis shows great potential for interoperative assessment of brain tissue during tumor surgery.
Intraoperative histology for brain tumors
In diseases such as interstitial lung diseases, immediate feedback on leukocyte ratios has potential to speed-up the diagnostic process and to reduce costs, workload and inter-observer variations. Studies have shown promising leukocyte identification performance on blood fractions, using third harmonic generation (THG) and multiphoton excited autofluorescence (MPEF) microscopy.
Immune cell differentiation in BAL fluids
Fast and reliable intraoperative pathological feedback using Higher harmonic generation microscopy on unprocessed tissue within minutes, is a promising tool to avoid a two-stage procedure and significantly reduce health care costs in patients undergoing a diagnostic hemithyroidectomy. Images generated show relevant morphological thyroid structures in good accordance with the histology images.
Avoid two-staged thyroid cancer surgery
Rapid and reliable histology assessment of tumor tissue can reduce operation time and potentially improve patient outcomes in lung cancer operations. THG/SHG/2PEF imaging in seconds of fresh, unprocesessed human lung tissue is a promising tool for clinical intraoperative assessment of lung tumor tissue. In this study, this digital histology is compared with standard FFPE HE stained histology images.
Showing histological hallmarks in lung tumor tissue
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