Abstract：
Fluorescence confocal microscopy (FCM) is an optical technique that uses laser light sources of different wavelengths to generate real-time images of fresh, unfixed tissue specimens. FCM allows histological evaluation of fresh tissue samples without the associated cryo artifacts after frozen sectioning. The aim of this study was to prospectively evaluate pediatric tumor specimens and assess their suitability for fresh tumor sampling. In addition, we aimed to determine whether tumor cell isolation for stable cell culture is still feasible after FCM imaging. Pediatric tumor specimens were imaged using FCM. Tumor viability and suitability for tissue sampling were evaluated and compared with H&E staining after paraffin embedding. In addition, FCM-processed and non-FCM-processed tissue samples were sent for tumor cell isolation to evaluate possible effects after FCM processing. When comparing estimated tumor cell viability using FCM and H&E, we found good to excellent correlating estimates (intraclass correlation coefficient = 0.891, p < 0.001), as well as substantial agreement in whether the tissue appeared adequate for fresh tissue collection (κ = 0.762, p < 0.001). After FCM, seven out of eight samples yielded passable cell cultures, compared to eight out of eight for non-FCM processed samples. Our study suggests that the use of FCM in tumor sampling can increase the yield of suitable fresh tumor samples by identifying viable tumor areas and ensuring that sufficient tissue remains for diagnosis. Our study also provides first evidence that the isolation and growth of tumor cells in culture are not compromised by the FCM technique.
摘要：
荧光共聚焦显微镜（FCM）是一种光学技术，它使用不同波长的激光光源来生成新鲜的实时图像，未固定的组织标本。FCM允许在冷冻切片后对新鲜组织样品进行组织学评估而没有相关的冷冻伪影。这项研究的目的是前瞻性评估小儿肿瘤标本，并评估其是否适合新鲜肿瘤采样。此外,我们的目的是确定FCM成像后肿瘤细胞分离进行稳定细胞培养是否仍然可行.使用FCM对小儿肿瘤标本进行成像。评估肿瘤活力和组织取样的适用性,并与石蜡包埋后的H&E染色进行比较。此外,将FCM处理的和非FCM处理的组织样品送去进行肿瘤细胞分离,以评估FCM处理后的可能效果。当使用FCM和H&E比较估计的肿瘤细胞活力时，我们发现了良好到极好的相关估计(组内相关系数=0.891，p<0.001)，以及该组织是否足以收集新鲜组织（κ=0.762，p<0.001）。在FCM之后，八个样本中有七个产生了可通过的细胞培养物，相比之下，非FCM处理样品的八分之八。我们的研究表明，在肿瘤采样中使用FCM可以通过确定可行的肿瘤区域并确保保留足够的组织来诊断，从而提高合适的新鲜肿瘤样品的产量。我们的研究还提供了初步证据，证明FCM技术不会损害培养物中肿瘤细胞的分离和生长。
