Abstract
.Accurate histopathological diagnosis is essential for facilitating the optimal surgical management of intracranial germinoma. Current intraoperative histological methods are time- and labor-intensive and often produce artifacts. Multiphoton microscopy (MPM) is a label-free imaging technique that can produce intraoperative histological images of fresh, unprocessed surgical specimens. We employ an MPM based on second-harmonic generation and two-photon excited fluorescence microscopy to image fresh, unfixed, and unstained human germinoma specimens. We show that label-free MPM is not only capable of identifying various cells in human germinoma tissue but also capable of revealing the characteristics of germinoma such as granuloma, stromal fibrosis, calcification, as well as the abnormal and uneven structures of blood vessels. In conjunction with custom-developed image-processing algorithms, MPM can further quantify and characterize the extent of stromal fibrosis and calcification. Our results provide insight into how MPM can deliver rapid diagnostic histological data that could inform the surgical management of intracranial germinoma.
Highlights
The accurate intraoperative histopathological diagnosis of intracranial germinoma is critical for determining the most appropriate surgical treatment, prognosis assessment, and predicting or evaluating a given patient’s response to a particular therapeutic approach.[1]
In the large-area images [Figs. 2(a)–2d)], the extracellular matrix, the cells, and the blood vessels that were the major brain tissue components emitted strong multiphoton signals, which might be explicitly distinguished in Multiphoton microscopy (MPM) images
Our results indicate that MPM technology can efficiently identify germinoma from the normal tissues
Summary
The accurate intraoperative histopathological diagnosis of intracranial germinoma is critical for determining the most appropriate surgical treatment, prognosis assessment, and predicting or evaluating a given patient’s response to a particular therapeutic approach.[1]. Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) is a more robust imaging tool that has attracted much attention since its inception.[6,7,8,9,10,11] This imaging modality is capable of producing images with a level of detail similar to that of standard histological procedures by detecting intrinsic fluorescence chromophores while demonstrating superior tissue penetration depth and reduced degrees of photobleaching and photodamage Owing to these significant advantages over confocal laser microscopy, MPM is widely applied in the investigation of healthy versus diseased brain tissues.[12,13,14,15,16,17]. To the best of our knowledge, no other study has been conducted around the use of MPM for examining human intracranial germinoma
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